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frameshifting rna pseudoknots: structure and mechanism.programmed ribosomal frameshifting (prf) is one of the multiple translational recoding processes that fundamentally alters triplet decoding of the messenger rna by the elongating ribosome. the ability of the ribosome to change translational reading frames in the -1 direction (-1 prf) is employed by many positive strand rna viruses, including economically important plant viruses and many human pathogens, such as retroviruses, e.g., hiv-1, and coronaviruses, e.g., the causative agent of severe acu ...200918621088
revealing unique properties of the ribosome using a network based analysis.the ribosome is a complex molecular machine that offers many potential sites for functional interference, therefore representing a major target for antibacterial drugs. the growing number of high-resolution structures of ribosomes from different organisms, in free form and in complex with various ligands, provides unique data for structural and comparative analyses of rna structures. we model the ribosome structure as a network, where nucleotides are represented as nodes and intermolecular inter ...200818625614
conserved discrimination against misacylated trnas by two mesophilic elongation factor tu orthologs.elongation factor tu (ef-tu) binds and loads elongating aminoacyl-trnas (aa-trnas) onto the ribosome for protein biosynthesis. many bacteria biosynthesize gln-trna (gln) and asn-trna (asn) by an indirect, two-step pathway that relies on the misacylated trnas glu-trna (gln) and asp-trna (asn) as intermediates. previous thermodynamic and experimental analyses have demonstrated that thermus thermophilus ef-tu does not bind asp-trna (asn) and predicted a similar discriminatory response against glu-t ...200818627126
the complete genome sequence of moorella thermoacetica (f. clostridium thermoaceticum).this paper describes the genome sequence of moorella thermoacetica (f. clostridium thermoaceticum), which is the model acetogenic bacterium that has been widely used for elucidating the wood-ljungdahl pathway of co and co(2) fixation. this pathway, which is also known as the reductive acetyl-coa pathway, allows acetogenic (often called homoacetogenic) bacteria to convert glucose stoichiometrically into 3 mol of acetate and to grow autotrophically using h(2) and co as electron donors and co(2) as ...200818631365
molecular mechanisms underlying the positive stringent response of the bacillus subtilis ilv-leu operon, involved in the biosynthesis of branched-chain amino acids.branched-chain amino acids are the most abundant amino acids in proteins. the bacillus subtilis ilv-leu operon is involved in the biosynthesis of branched-chain amino acids. this operon exhibits a rela-dependent positive stringent response to amino acid starvation. we investigated this positive stringent response upon lysine starvation as well as decoyinine treatment. deletion analysis involving various lacz fusions revealed two molecular mechanisms underlying the positive stringent response of ...200818641142
visualization of the eef2-80s ribosome transition-state complex by cryo-electron microscopy.in an attempt to understand ribosome-induced gtp hydrolysis on eef2, we determined a 12.6-a cryo-electron microscopy reconstruction of the eef2-bound 80s ribosome in the presence of aluminum tetrafluoride and gdp, with aluminum tetrafluoride mimicking the gamma-phosphate during hydrolysis. this is the first visualization of a structure representing a transition-state complex on the ribosome. tight interactions are observed between the factor's g domain and the large ribosomal subunit, as well as ...200818644383
erythromycin-induced ribosome stalling and rnase j1-mediated mrna processing in bacillus subtilis.summary: addition of erythromycin (em) to a bacillus subtilis strain carrying the ermc gene results in ribosome stalling in the ermc leader peptide coding sequence. using deltaermc, a deletion derivative of ermc that specifies the 254 nucleotide deltaermc mrna, we showed previously that ribosome stalling is concomitant with processing of deltaermc mrna, generating a 209 nucleotide rna whose 5' end maps to codon 5 of the deltaermc coding sequence. here we probed for peptidyl-trna to show that rib ...200818647167
structural frameworks for considering microbial protein- and nucleic acid-dependent motor atpases.many fundamental cellular processes depend on enzymes that utilize chemical energy to catalyse unfavourable reactions. certain classes of atpases provide a particularly vivid example of the process of energy conversion, employing cycles of nucleotide turnover to move and/or rearrange biological polymers such as proteins and nucleic acids. four well-characterized classes of atp-dependent protein/nucleic acid translocases and remodelling factors are found in all three domains of life (bacteria, ar ...200818647240
the highest affinity binding site of small protein b on transfer messenger rna is outside the trna domain.eubacterial ribosomes stalled on defective mrnas are released through a mechanism referred to as trans-translation, depending on the coordinated actions of small protein b (smpb) and transfer messenger rna (tmrna). a series of tmrna variants with deletions in each structural domain were produced. their structures were monitored by enzymatic and chemical probes in vitro, in the presence and absence of smpb. dissociation constants between these rnas and smpb from aquifex aeolicus were derived by s ...200818648069
a new regulatory circuit in ribosomal protein operons: s2-mediated control of the rpsb-tsf expression in vivo.autogenous regulation is a general strategy of balancing ribosomal protein synthesis in bacteria. control mechanisms have been studied in detail for most of ribosomal protein operons, except for rpsb-tsf encoding essential r-protein s2 and elongation factor ts, where even the promoter has remained unknown. by using single-copy translational fusions with the chromosomal lacz gene and western-blot analysis, we demonstrate here that s2 serves as a negative regulator of both rpsb and tsf expression ...200818648071
crenarchaeal arginine decarboxylase evolved from an s-adenosylmethionine decarboxylase enzyme.the crenarchaeon sulfolobus solfataricus uses arginine to produce putrescine for polyamine biosynthesis. however, genome sequences from s. solfataricus and most crenarchaea have no known homologs of the previously characterized pyridoxal 5'-phosphate or pyruvoyl-dependent arginine decarboxylases that catalyze the first step in this pathway. instead they have two paralogs of the s-adenosylmethionine decarboxylase (adometdc). the gene at locus sso0585 produces an adometdc enzyme, whereas the gene ...200818650422
structure and mechanistic implications of a uroporphyrinogen iii synthase-product complex.uroporphyrinogen iii synthase (u3s) catalyzes the asymmetrical cyclization of a linear tetrapyrrole to form the physiologically relevant uroporphyrinogen iii (uro'gen iii) isomer during heme biosynthesis. here, we report four apoenzyme and one product complex crystal structures of the thermus thermophilus (hb27) u3s protein. the overlay of eight crystallographically unique u3s molecules reveals a huge range of conformational flexibility, including a "closed" product complex. the product, uro'gen ...200818651750
distinct double- and single-stranded dna binding of e. coli replicative dna polymerase iii alpha subunit.the alpha subunit of the replicative dna polymerase iii of escherichia coli is the active polymerase of the 10-subunit bacterial replicase. the c-terminal region of the alpha subunit is predicted to contain an oligonucleotide binding (ob-fold) domain. in a series of optical tweezers experiments, the alpha subunit is shown to have an affinity for both double- and single-stranded dna, in distinct subdomains of the protein. the portion of the protein that binds to double-stranded dna stabilizes the ...200818652472
origin of the nucleus and ran-dependent transport to safeguard ribosome biogenesis in a chimeric cell.the origin of the nucleus is a central problem about the origin of eukaryotes. the common ancestry of nuclear pore complexes (npc) and vesicle coating complexes indicates that the nucleus evolved via the modification of a pre-existing endomembrane system. such an autogenous scenario is cell biologically feasible, but it is not clear what were the selective or neutral mechanisms that had led to the origin of the nuclear compartment.200818652645
a unique conformation of the anticodon stem-loop is associated with the capacity of trnafmet to initiate protein synthesis.in all organisms, translational initiation takes place on the small ribosomal subunit and two classes of methionine trna are present. the initiator is used exclusively for initiation of protein synthesis while the elongator is used for inserting methionine internally in the nascent polypeptide chain. the crystal structure of escherichia coli initiator trna(f)(met) has been solved at 3.1 a resolution. the anticodon region is well-defined and reveals a unique structure, which has not been describe ...200818653533
recr forms a ring-like tetramer that encircles dsdna by forming a complex with recf.in the recfor pathway, the recf and recr proteins form a complex that binds to dna and exerts multiple functions, including directing the loading of reca onto single-stranded (ss) dna regions near double-stranded (ds) dna-ssdna junctions and preventing it from forming a filament beyond the ssdna region. however, neither the structure of the recfr complex nor its dna-binding mechanism was previously identified. here, size-exclusion chromatography and small-angle x-ray scattering data indicate tha ...200818658243
crystal structure of the thermus thermophilus 16 s rrna methyltransferase rsmc in complex with cofactor and substrate guanosine.post-transcriptional modification is a ubiquitous feature of ribosomal rna in all kingdoms of life. modified nucleotides are generally clustered in functionally important regions of the ribosome, but the functional contribution to protein synthesis is not well understood. here we describe high resolution crystal structures for the n(2)-guanine methyltransferase rsmc that modifies residue g1207 in 16 s rrna near the decoding site of the 30 s ribosomal subunit. rsmc is a class i s-adenosyl-l-methi ...200818667428
the rna acetyltransferase driven by atp hydrolysis synthesizes n4-acetylcytidine of trna anticodon.the wobble base of escherichia coli elongator trna(met) is modified to n(4)-acetylcytidine (ac(4)c), which is thought to ensure the precise recognition of the aug codon by preventing misreading of near-cognate aua codon. by employing genome-wide screen of uncharacterized genes in escherichia coli ('ribonucleome analysis'), we found the ypfi gene, which we named tmca (trna(met) cytidine acetyltransferase), to be responsible for ac(4)c formation. tmca is an enzyme that contains a walker-type atpas ...200818668122
ppargamma and proline oxidase in cancer.proline is metabolized by its own specialized enzymes with their own tissue and subcellular localizations and mechanisms of regulation. the central enzyme in this metabolic system is proline oxidase, a flavin adenine dinucleotide-containing enzyme which is tightly bound to mitochondrial inner membranes. the electrons from proline can be used to generate atp or can directly reduce oxygen to form superoxide. although proline may be derived from the diet and biosynthesized endogenously, an importan ...200818670615
thermodynamic redox behavior of the heme centers in a-type heme-copper oxygen reductases: comparison between the two subfamilies.the study of the thermodynamic redox behavior of the hemes from two members of the a family of heme-copper oxygen reductases, paracoccus denitrificans aa3 (a1 subfamily) and rhodothermus marinus caa3 (a2 subfamily) enzymes, is presented. at different ph values, midpoint reduction potentials and interaction potentials were obtained in the framework of a pairwise model for two interacting redox centers. in both enzymes, the hemes have different reduction potentials. for the a1-type enzyme, it was ...200818676644
deciphering the genetic determinants for aerobic nicotinic acid degradation: the nic cluster from pseudomonas putida kt2440.the aerobic catabolism of nicotinic acid (na) is considered a model system for degradation of n-heterocyclic aromatic compounds, some of which are major environmental pollutants; however, the complete set of genes as well as the structural-functional relationships of most of the enzymes involved in this process are still unknown. we have characterized a gene cluster (nic genes) from pseudomonas putida kt2440 responsible for the aerobic na degradation in this bacterium and when expressed in heter ...200818678916
cloning, expression, purification and preliminary crystallographic analysis of the rnase hi domain of the mycobacterium tuberculosis protein rv2228c as a maltose-binding protein fusion.the predicted ribonuclease (rnase) hi domain of the open reading frame rv2228c from mycobacterium tuberculosis has been cloned as a hexahistidine fusion and a maltose-binding protein (mbp) fusion. expression was only observed for the mbp-fusion protein, which was purified using amylose affinity chromatography and gel filtration. the rnase hi domain could be cleaved from the mbp-fusion protein by factor xa digestion, but was very unstable. in contrast, the fusion protein was stable, could be obta ...200818678948
structure of the e. coli dna glycosylase alka bound to the ends of duplex dna: a system for the structure determination of lesion-containing dna.the constant attack on dna by endogenous and exogenous agents gives rise to nucleobase modifications that cause mutations, which can lead to cancer. visualizing the effects of these lesions on the structure of duplex dna is key to understanding their biologic consequences. the most definitive method of obtaining such structures, x-ray crystallography, is troublesome to employ owing to the difficulty of obtaining diffraction-quality crystals of dna. here, we present a crystallization system that ...200818682218
hyperthermophilic aquifex aeolicus initiates primer synthesis on a limited set of trinucleotides comprised of cytosines and guanines.the placement of the extreme thermophile aquifex aeolicus in the bacterial phylogenetic tree has evoked much controversy. we investigated whether adaptations for growth at high temperatures would alter a key functional component of the replication machinery, specifically dnag primase. although the structure of bacterial primases is conserved, the trinucleotide initiation specificity for a. aeolicus was hypothesized to differ from other microbes as an adaptation to a geothermal milieu. to determi ...200818684998
conserved motifs in both cpsf73 and cpsf100 are required to assemble the active endonuclease for histone mrna 3'-end maturation.in eukaryotes, the process of messenger rna 3'-end formation involves endonucleolytic cleavage of the transcript followed by synthesis of the poly(a) tail. the complex machinery involved in this maturation process contains two proteins of the metallo-beta-lactamase (mbl) superfamily, the 73 and 100 kda subunits of the cleavage and polyadenylation specificity factor (cpsf). by using an in vitro system to assess point mutations in these two mammalian proteins, we found that conserved residues from ...200818688255
the growth-promoting and stress response activities of the bacillus subtilis gtp binding protein obg are separable by mutation.bacillus subtilis obg is a ribosome-associating gtp binding protein that is needed for growth, sporulation, and induction of the bacterium's general stress regulon (gsr). it is unclear whether the roles of obg in sporulation and stress responsiveness are direct or a secondary effect of its growth-promoting functions. the present work addresses this question by an analysis of two obg alleles whose phenotypes argue for direct roles for obg in each process. the first allele [obg(g92d)] encodes a mi ...200818689482
insights into the replisome from the structure of a ternary complex of the dna polymerase iii alpha-subunit.the crystal structure of the catalytic alpha-subunit of the dna polymerase iii (pol iiialpha) holoenzyme bound to primer-template dna and an incoming deoxy-nucleoside 5'-triphosphate has been determined at 4.6-a resolution. the polymerase interacts with the sugar-phosphate backbone of the dna across its minor groove, which is made possible by significant movements of the thumb, finger, and beta-binding domains relative to their orientations in the unliganded polymerase structure. additionally, t ...200818691598
crystal structure of type 2 isopentenyl diphosphate isomerase from thermus thermophilus in complex with inorganic pyrophosphate.the n-terminal region is stabilized in the crystal structure of thermus thermophilus type 2 isopentenyl diphosphate isomerase in complex with inorganic pyrophosphate, providing new insights about the active site and the catalytic mechanism of the enzyme. the pp i moiety is located near the conserved residues, h10, r97, h152, q157, e158, and w219, and the flavin cofactor. the putative active site of isopentenyl diphosphate isomerase 2 provides interactions for stabilizing a carbocationic intermed ...200818693754
crystal structures of nadh:fmn oxidoreductase (emob) at different stages of catalysis.edta has become a major organic pollutant in the environment because of its extreme usage and resistance to biodegradation. recently, two critical enzymes, edta monooxygenase (emoa) and nadh:fmn oxidoreductase (emob), belonging to the newly established two-component flavin-diffusible monooxygenase family, were identified in the edta degradation pathway in mesorhizobium sp. bnc1. emoa is an fmnh2-dependent enzyme that requires emob to provide fmnh2 for the conversion of edta to ethylenediaminedia ...200818701448
mitochondrial nadh fluorescence is enhanced by complex i binding.mitochondrial nadh fluorescence has been a useful tool in evaluating mitochondrial energetics both in vitro and in vivo. mitochondrial nadh fluorescence is enhanced several-fold in the matrix through extended fluorescence lifetimes (efl). however, the actual binding sites responsible for nadh efl are unknown. we tested the hypothesis that nadh binding to complex i is a significant source of mitochondrial nadh fluorescence enhancement. to test this hypothesis, the effect of complex i binding on n ...200818702505
molecular dynamics guided study of salt bridge length dependence in both fluorinated and non-fluorinated parallel dimeric coiled-coils.the alpha-helical coiled-coil is one of the most common oligomerization motifs found in both native and engineered proteins. to better understand the stability and dynamics of the coiled-coil motifs, including those modified by fluorination, several fluorinated and nonfluorinated parallel dimeric coiled-coil protein structures were designed and modeled. we also attempt to investigate how changing the length and geometry of the important stabilizing salt bridges influences the coiled-coil protein ...200918704948
genetic identification of yeast 18s rrna residues required for efficient recruitment of initiator trna(met) and aug selection.high-resolution structures of bacterial 70s ribosomes have provided atomic details about mrna and trna binding to the decoding center during elongation, but such information is lacking for preinitiation complexes (pics). we identified residues in yeast 18s rrna critical in vivo for recruiting methionyl trna(i)(met) to 40s subunits during initiation by isolating mutations that derepress gcn4 mrna translation. several such gcd(-) mutations alter the a928:u1389 base pair in helix 28 (h28) and allow ...200818708582
slr1923 of synechocystis sp. pcc6803 is essential for conversion of 3,8-divinyl(proto)chlorophyll(ide) to 3-monovinyl(proto)chlorophyll(ide).the deduced amino acid sequence of an slr1923 gene of synechocystis sp. pcc6803 is homologous to archaean f(420)h(2) dehydrogenase, which acts as a soluble subcomplex of reduced nicotinamide adenine dinucleotide dehydrogenase complex i. in this study, the gene was inactivated and characteristics of the mutant were analyzed. the mutant grew slower than the wild type under 100 microe m(-2) s(-1) but did not grow under high light intensity (300 microe m(-2) s(-1)). the cellular content of chlorophy ...200818715956
s-adenosyl-l-methionine hydrolase (adenosine-forming), a conserved bacterial and archaeal protein related to sam-dependent halogenases. 200818720493
purification and characterization of the bacterial udp-glcnac:undecaprenyl-phosphate glcnac-1-phosphate transferase weca.to date, the structural and functional characterization of proteins belonging to the polyprenyl-phosphate n-acetylhexosamine-1-phosphate transferase superfamily has been relentlessly held back by problems encountered with their overexpression and purification. in the present work and for the first time, the integral membrane protein weca that catalyzes the transfer of the glcnac-1-phosphate moiety from udp-glcnac onto the carrier lipid undecaprenyl phosphate, yielding undecaprenyl-pyrophosphoryl ...200818723618
the rate and character of spontaneous mutation in thermus thermophilus.selection of spontaneous, loss-of-function mutations at two chromosomal loci (pyrf and pyre) enabled the first molecular-level analysis of replication fidelity in the extremely thermophilic bacterium thermus thermophilus. two different methods yielded similar mutation rates, and mutational spectra determined by sequencing of independent mutants revealed a variety of replication errors distributed throughout the target genes. the genomic mutation rate estimated from these targets, 0.00097 +/- 0.0 ...200818723895
alignment of protein structures in the presence of domain motions.structural alignment is an important step in protein comparison. well-established methods exist for solving this problem under the assumption that the structures under comparison are considered as rigid bodies. however, proteins are flexible entities often undergoing movements that alter the positions of domains or subdomains with respect to each other. such movements can impede the identification of structural equivalences when rigid aligners are used.200818727838
one heme, diverse functions: using biosynthetic myoglobin models to gain insights into heme-copper oxidases and nitric oxide reductases. 200818729107
the d subunit plays a central role in human vacuolar h(+)-atpases.the multi-subunit vacuolar-type h(+)-atpase consists of a v(1) domain (a-h subunits) catalyzing atp hydrolysis and a v(0) domain (a, c, c', c", d, e) responsible for h(+) translocation. the mammalian v(0) d subunit is one of the least-well characterized, and its function and position within the pump are still unclear. it has two different forms encoded by separate genes, d1 being ubiquitous while d2 is predominantly expressed at the cell surface in kidney and osteoclast. to determine whether it ...200818752060
electron and proton transfer in the ba(3) oxidase from thermus thermophilus.the ba(3)-type cytochrome c oxidase from thermus thermophilus is phylogenetically very distant from the aa(3)-type cytochrome c oxidases. nevertheless, both types of oxidases have the same number of redox-active metal sites and the reduction of o(2) to water is catalysed at a haem a(3)-cu(b) catalytic site. the three-dimensional structure of the ba(3) oxidase reveals three possible proton-conducting pathways showing very low homology compared to those of the mitochondrial, rhodobacter sphaeroide ...200818752061
structure of the guide-strand-containing argonaute silencing complex.the slicer activity of the rna-induced silencing complex is associated with argonaute, the rnase h-like piwi domain of which catalyses guide-strand-mediated sequence-specific cleavage of target messenger rna. here we report on the crystal structure of thermus thermophilus argonaute bound to a 5'-phosphorylated 21-base dna guide strand, thereby identifying the nucleic-acid-binding channel positioned between the paz- and piwi-containing lobes, as well as the pivot-like conformational changes assoc ...200818754009
ybea is the m3psi methyltransferase rlmh that targets nucleotide 1915 in 23s rrna.pseudouridines in the stable rnas of bacteria are seldom subjected to further modification. there are 11 pseudouridine (psi) sites in escherichia coli rrna, and further modification is found only at psi1915 in 23s rrna, where the n-3 position of the base becomes methylated. here, we report the identity of the e. coli methyltransferase that specifically catalyzes methyl group addition to form m(3)psi1915. analyses of e. coli rrnas using maldi mass spectrometry showed that inactivation of the ybea ...200818755835
the oxazolidinone antibiotics perturb the ribosomal peptidyl-transferase center and effect trna positioning.the oxazolidinones represent the first new class of antibiotics to enter into clinical usage within the past 30 years, but their binding site and mechanism of action has not been fully characterized. we have determined the crystal structure of the oxazolidinone linezolid bound to the deinococcus radiodurans 50s ribosomal subunit. linezolid binds in the a site pocket at the peptidyltransferase center of the ribosome overlapping the aminoacyl moiety of an a-site bound trna as well as many clinical ...200818757750
mechanism of cu(a) assembly.copper is essential for proper functioning of cytochrome c oxidases, and therefore for cellular respiration in eukaryotes and many bacteria. here we show that a new periplasmic protein (pcu(a)c) selectively inserts cu(i) ions into subunit ii of thermus thermophilus ba(3) oxidase to generate a native cu(a) site. the purported metallochaperone sco1 is unable to deliver copper ions; instead, it works as a thiol-disulfide reductase to maintain the correct oxidation state of the cu(a) cysteine ligand ...200818758441
crystal structure of escherichia coli rnk, a new rna polymerase-interacting protein.sequence-based searches identified a new family of genes in proteobacteria, named rnk, which shares high sequence similarity with the c-terminal domains of the gre factors (grea and greb) and the thermus/deinococcus anti-gre factor gfh1. we solved the x-ray crystal structure of escherichia coli regulator of nucleoside kinase (rnk) at 1.9 a resolution using the anomalous signal from the native protein. the rnk structure strikingly resembles those of e. coli grea and greb and thermus gfh1, all of ...200818760284
redox regulation of protein folding in the mitochondrial intermembrane space.protein translocation pathways to the mitochondrial matrix and inner membrane have been well characterized. however, translocation into the intermembrane space, which was thought to be simply a modification of the traditional translocation pathways, is complex. the mechanism by which a subset of intermembrane space proteins, those with disulfide bonds, are translocated has been largely unknown until recently. specifically, the intermembrane space proteins with disulfide bonds are imported via th ...200818761382
redox regulation of protein folding in the mitochondrial intermembrane space.protein translocation pathways to the mitochondrial matrix and inner membrane have been well characterized. however, translocation into the intermembrane space, which was thought to be simply a modification of the traditional translocation pathways, is complex. the mechanism by which a subset of intermembrane space proteins, those with disulfide bonds, are translocated has been largely unknown until recently. specifically, the intermembrane space proteins with disulfide bonds are imported via th ...200818761382
methanogen homoaconitase catalyzes both hydrolyase reactions in coenzyme b biosynthesis.homoaconitase enzymes catalyze hydrolyase reactions in the alpha-aminoadipate pathway for lysine biosynthesis or the 2-oxosuberate pathway for methanogenic coenzyme b biosynthesis. despite the homology of this iron-sulfur protein to aconitase, previously studied homoaconitases catalyze only the hydration of cis-homoaconitate to form homoisocitrate rather than the complete isomerization of homocitrate to homoisocitrate. the mj1003 and mj1271 proteins from the methanogen methanocaldococcus jannasc ...200818765671
transplantation of a tyrosine editing domain into a tyrosyl-trna synthetase variant enhances its specificity for a tyrosine analog.to guarantee specific trna and amino acid pairing, several aminoacyl-trna synthetases correct aminoacylation errors by deacylating or "editing" misaminoacylated trna. a previously developed variant of escherichia coli tyrosyl-trna synthetase (iodotyrrs) esterifies or "charges" trna(tyr) with a nonnatural amino acid, 3-iodo-l-tyrosine, and with l-tyrosine less efficiently. in the present study, the editing domain of phenylalanyl-trna synthetase (phers) was transplanted into iodotyrrs to edit tyro ...200818765802
development of trna synthetases and connection to genetic code and disease.the genetic code is established by the aminoacylation reactions of aminoacyl trna synthetases, where amino acids are matched with triplet anticodons imbedded in the cognate trnas. the code established in this way is so robust that it gave birth to the entire tree of life. the trna synthetases are organized into two classes, based on their active site architectures. the details of this organization, and other considerations, suggest how the synthetases evolved by gene duplications, and how early ...200818765819
enzymes used in molecular biology: a useful guide.since molecular cloning has become routine laboratory technique, manufacturers offer countless sources of enzymes to generate and manipulate nucleic acids. thus, selecting the appropriate enzyme for a specific task may seem difficult to the novice. this review aims at providing the readers with some cues for understanding the function and specificities of the different sources of polymerases, ligases, nucleases, phosphatases, methylases, and topoisomerases used for molecular cloning. we provide ...200818766469
crystal structure of a sulfur carrier protein complex found in the cysteine biosynthetic pathway of mycobacterium tuberculosis.the structure of the protein complex cysm-cyso from a new cysteine biosynthetic pathway found in the h37rv strain of mycobacterium tuberculosis has been determined at 1.53 a resolution. cysm (rv1336) is a plp-containing beta-replacement enzyme and cyso (rv1335) is a sulfur carrier protein with a ubiquitin-like fold. cysm catalyzes the replacement of the acetyl group of o-acetylserine by cyso thiocarboxylate to generate a protein-bound cysteine that is released in a subsequent proteolysis reactio ...200818771296
full-length escherichia coli seca dimerizes in a closed conformation in solution as determined by cryo-electron microscopy.seca is an obligatory component of the escherichia coli general secretion pathway. however, the oligomeric structure of seca and seca conformational changes during translocation processes are still unclear. here we obtained the three-dimensional structure of e. coli wild-type full-length seca in solution by single particle cryo-electron microscopy and determined its oligomeric organization. in this structure, seca occurs as a dimer in which the two protomers are arranged in an antiparallel mode, ...200818772144
mechanism of the chemical step for the guanosine triphosphate (gtp) hydrolysis catalyzed by elongation factor tu.elongation factor tu (ef-tu), the protein responsible for delivering aminoacyl-trnas (aa-trnas) to ribosomal a site during translation, belongs to the group of guanosine-nucleotide (gtp/gdp) binding proteins. its active 'on'-state corresponds to the gtp-bound form, while the inactive 'off'-state corresponds to the gdp-bound form. in this work we focus on the chemical step, gtp+h(2)o-->gdp+pi, of the hydrolysis mechanism. we apply molecular modeling tools including molecular dynamics simulations ...200818773979
inhibition of a transcriptional pause by rna anchoring to rna polymerase.we describe a mechanism by which nascent rna inhibits transcriptional pausing. putl rna of bacteriophage hk022 suppresses transcription termination at downstream terminators and pausing within a nearby u-rich sequence. in vitro transcription and footprinting assays reveal that this pausing results from backtracking of rna polymerase and that binding of nascent putl rna to polymerase limits backtracking by restricting re-entry of the transcript into the rna exit channel. the restriction is local ...200818775328
genetic and biochemical analysis of yeast and human cap trimethylguanosine synthase: functional overlap of 2,2,7-trimethylguanosine caps, small nuclear ribonucleoprotein components, pre-mrna splicing factors, and rna decay pathways.trimethylguanosine synthase (tgs1) is the enzyme that converts standard m(7)g caps to the 2,2,7-trimethylguanosine (tmg) caps characteristic of spliceosomal small nuclear rnas. fungi and mammalian somatic cells are able to grow in the absence of tgs1 and tmg caps, suggesting that an essential function of the tmg cap might be obscured by functional redundancy. a systematic screen in budding yeast identified nonessential genes that, when deleted, caused synthetic growth defects with tgs1delta. the ...200818775984
a novel mutator of escherichia coli carrying a defect in the dgt gene, encoding a dgtp triphosphohydrolase.a novel mutator locus in escherichia coli was identified from a collection of random transposon insertion mutants. several mutators in this collection were found to have an insertion in the dgt gene, encoding a previously characterized dgtp triphosphohydrolase. the mutator activity of the dgt mutants displays an unusual specificity. among the six possible base pair substitutions in a lacz reversion system, the g.c-->c.g transversion and a.t-->g.c transition are strongly enhanced (10- to 50-fold) ...200818776019
editing of misaligned 3'-termini by an intrinsic 3'-5' exonuclease activity residing in the php domain of a family x dna polymerase.bacillus subtilis gene yshc encodes a family x dna polymerase (polx(bs)), whose biochemical features suggest that it plays a role during dna repair processes. here, we show that, in addition to the polymerization activity, polx(bs) possesses an intrinsic 3'-5' exonuclease activity specialized in resecting unannealed 3'-termini in a gapped dna substrate. biochemical analysis of a polx(bs) deletion mutant lacking the c-terminal polymerase histidinol phosphatase (php) domain, present in most of the ...200818776221
structural biology of riboswitch-mediated gene regulation and argonaute-mediated gene silencing. 200818776223
characterization of a unique clpb protein of mycoplasma pneumoniae and its impact on growth.mycoplasma pneumoniae accounts for 20 to 30% of all community-acquired pneumonia and has been associated with other airway pathologies, including asthma, and a range of extrapulmonary manifestations. although the entire genomic sequence of m. pneumoniae has been completed, the functions of many of these genes in mycoplasma physiology are unknown. in this study, we focused on clpb, a well-known heat shock gene in other bacteria, to examine its role in mycoplasma growth. transcriptional and transl ...200818779336
stabilization of alpha-chymotrypsin upon pegylation correlates with reduced structural dynamics.protein stability remains one of the main factors limiting the realization of the full potential of protein therapeutics. poly(ethylene glycol) (peg) conjugation to proteins has evolved into an important tool to overcome instability issues associated with proteins. the observed increase in thermodynamic stability of several proteins upon pegylation has been hypothesized to arise from reduced protein structural dynamics, although experimental evidence for this hypothesis is currently missing. to ...200818781698
synthesis and characterization of new piperazine-type inhibitors for mitochondrial nadh-ubiquinone oxidoreductase (complex i).the mode of action of deltalac-acetogenins, strong inhibitors of bovine heart mitochondrial complex i, is different from that of traditional inhibitors such as rotenone and piericidin a [murai, m., et al. (2007) biochemistry 46 , 6409-6416]. as further exploration of these unique inhibitors might provide new insights into the terminal electron transfer step of complex i, we drastically modified the structure of deltalac-acetogenins and characterized their inhibitory action. in particular, on the ...200818781777
the putative rnase p motif in the dead box helicase hera is dispensable for efficient interaction with rna and helicase activity.dead box helicases use the energy of atp hydrolysis to remodel rna structures or rna/protein complexes. they share a common helicase core with conserved signature motifs, and additional domains may confer substrate specificity. identification of a specific substrate is crucial towards understanding the physiological role of a helicase. rna binding and atpase stimulation are necessary, but not sufficient criteria for a bona fide helicase substrate. here, we report single molecule fret experiments ...200818782831
the human mitochondrial ribosome recycling factor is essential for cell viability.the molecular mechanism of human mitochondrial translation has yet to be fully described. we are particularly interested in understanding the process of translational termination and ribosome recycling in the mitochondrion. several candidates have been implicated, for which subcellular localization and characterization have not been reported. here, we show that the putative mitochondrial recycling factor, mtrrf, is indeed a mitochondrial protein. expression of human mtrrf in fission yeast devoid ...200818782833
concurrent nucleation of 16s folding and induced fit in 30s ribosome assembly.rapidly growing cells produce thousands of new ribosomes each minute, in a tightly regulated process that is essential to cell growth. how the escherichia coli 16s ribosomal rna and the 20 proteins that make up the 30s ribosomal subunit can assemble correctly in a few minutes remains a challenging problem, partly because of the lack of real-time data on the earliest stages of assembly. by providing snapshots of individual rna and protein interactions as they emerge in real time, here we show tha ...200818784650
insights into the mode of action of a putative zinc transporter czrb in thermus thermophilus.the crystal structures of the cytoplasmic domain of the putative zinc transporter czrb in the apo and zinc-bound forms reported herein are consistent with the protein functioning in vivo as a homodimer. nmr, x-ray scattering, and size-exclusion chromatography provide support for dimer formation. full-length variants of czrb in the apo and zinc-loaded states were generated by homology modeling with the zn2+/h+ antiporter yiip. the model suggests a way in which zinc binding to the cytoplasmic frag ...200818786400
rifamycins do not function by allosteric modulation of binding of mg2+ to the rna polymerase active center.rifamycin antibacterial agents inhibit bacterial rna polymerase (rnap) by binding to a site adjacent to the rnap active center and preventing synthesis of rna products >2-3 nt in length. recently, artsimovitch et al. [(2005) cell 122:351-363] proposed that rifamycins function by allosteric modulation of binding of mg(2+) to the rnap active center and presented three lines of biochemical evidence consistent with this proposal. here, we show that rifamycins do not affect the affinity of binding of ...200818787125
mass spectrometry profiles superoxide-induced intramolecular disulfide in the fmn-binding subunit of mitochondrial complex i.protein thiols with regulatory functions play a critical role in maintaining the homeostasis of the redox state in mitochondria. one major host of regulatory cysteines in mitochondria is complex i, with the thiols primarily located on its 51 kda fmn-binding subunit. in response to oxidative stress, these thiols are expected to form intramolecular disulfide bridges as one of their oxidative post-translational modifications. here, to test this hypothesis and gain insights into the molecular patter ...200818789718
the mycobacterium tuberculosis mep (2c-methyl-d-erythritol 4-phosphate) pathway as a new drug target.tuberculosis (tb) is still a major public health problem, compounded by the human immunodeficiency virus (hiv)-tb co-infection and recent emergence of multidrug-resistant (mdr) and extensively drug resistant (xdr)-tb. novel anti-tb drugs are urgently required. in this context, the 2c-methyl-d-erythritol 4-phosphate (mep) pathway of mycobacterium tuberculosis has drawn attention; it is one of several pathways vital for m. tuberculosis viability and the human host lacks homologous enzymes. thus, t ...200918793870
the mycobacterium tuberculosis mep (2c-methyl-d-erythritol 4-phosphate) pathway as a new drug target.tuberculosis (tb) is still a major public health problem, compounded by the human immunodeficiency virus (hiv)-tb co-infection and recent emergence of multidrug-resistant (mdr) and extensively drug resistant (xdr)-tb. novel anti-tb drugs are urgently required. in this context, the 2c-methyl-d-erythritol 4-phosphate (mep) pathway of mycobacterium tuberculosis has drawn attention; it is one of several pathways vital for m. tuberculosis viability and the human host lacks homologous enzymes. thus, t ...200918793870
genome signature analysis of thermal virus metagenomes reveals archaea and thermophilic signatures.metagenomic analysis provides a rich source of biological information for otherwise intractable viral communities. however, study of viral metagenomes has been hampered by its nearly complete reliance on blast algorithms for identification of dna sequences. we sought to develop algorithms for examination of viral metagenomes to identify the origin of sequences independent of blast algorithms. we chose viral metagenomes obtained from two hot springs, bear paw and octopus, in yellowstone national ...200818798991
mutations in conserved helix 69 of 23s rrna of thermus thermophilus that affect capreomycin resistance but not posttranscriptional modifications.translocation during the elongation phase of protein synthesis involves the relative movement of the 30s and 50s ribosomal subunits. this movement is the target of tuberactinomycin antibiotics. here, we describe the isolation and characterization of mutants of thermus thermophilus selected for resistance to the tuberactinomycin antibiotic capreomycin. two base substitutions, a1913u and mu1915g, and a single base deletion, deltamu1915, were identified in helix 69 of 23s rrna, a structural element ...200818805973
mechanism of 4-nitrophenol oxidation in rhodococcus sp. strain pn1: characterization of the two-component 4-nitrophenol hydroxylase and regulation of its expression.4-nitrophenol (4-np) is a toxic product of the hydrolysis of organophosphorus pesticides such as parathion in soil. rhodococcus sp. strain pn1 degrades 4-np via 4-nitrocatechol (4-nc) for use as the sole carbon, nitrogen, and energy source. a 5-kb ecori dna fragment previously cloned from pn1 contained a gene cluster (nphra1a2) involved in 4-np oxidation. from sequence analysis, this gene cluster is expected to encode an arac/xyls family regulatory protein (nphr) and a two-component 4-np hydroxy ...200818805976
the metabolism of proline as microenvironmental stress substrate.proline, a unique proteogenic secondary amino acid, has its own metabolic system with special features. recent findings defining the regulation of this system led us to propose that proline is a stress substrate in the microenvironment of inflammation and tumorigenesis. the criteria for proline as a stress substrate are: 1) the enzymes utilizing proline respond to stress signaling; 2) there is a large, mobilizable pool of proline; and 3) the metabolism of proline serves special stress functions. ...200818806116
structural analysis of fad synthetase from corynebacterium ammoniagenes.the prokaryotic fad synthetase family - a group of bifunctional enzymes that catalyse riboflavin phosphorylation and fmn adenylylation within a single polypeptide chain- was analysed in terms of sequence and structure.200818811972
endosymbiont gene functions impaired and rescued by polymerase infidelity at poly(a) tracts.among host-dependent bacteria that have evolved by extreme reductive genome evolution, long-term bacterial endosymbionts of insects have the smallest (160-790 kb) and most a + t-rich (>70%) bacterial genomes known to date. these genomes are riddled with poly(a) tracts, and 5-50% of genes contain tracts of 10 as or more. here, we demonstrate transcriptional slippage at poly(a) tracts within genes of buchnera aphidicola associated with aphids and blochmannia pennsylvanicus associated with ants. se ...200818815381
molecular characterization of organelle-type nudix hydrolases in arabidopsis.nudix (for nucleoside diphosphates linked to some moiety x) hydrolases act to hydrolyze ribonucleoside and deoxyribonucleoside triphosphates, nucleotide sugars, coenzymes, or dinucleoside polyphosphates. arabidopsis (arabidopsis thaliana) contains 27 genes encoding nudix hydrolase homologues (atnudx1 to -27) with a predicted distribution in the cytosol, mitochondria, and chloroplasts. previously, cytosolic nudix hydrolases (atnudx1 to -11 and -25) were characterized. here, we conducted a charact ...200818815383
protein co-evolution, co-adaptation and interactions.co-evolution has an important function in the evolution of species and it is clearly manifested in certain scenarios such as host-parasite and predator-prey interactions, symbiosis and mutualism. the extrapolation of the concepts and methodologies developed for the study of species co-evolution at the molecular level has prompted the development of a variety of computational methods able to predict protein interactions through the characteristics of co-evolution. particularly successful have bee ...200818818697
yeast ribosomal protein l10 helps coordinate trna movement through the large subunit.yeast ribosomal protein l10 (e. coli l16) is located at the center of a topological nexus that connects many functional regions of the large subunit. this essential protein has previously been implicated in processes as diverse as ribosome biogenesis, translational fidelity and mrna stability. here, the inability to maintain the yeast killer virus was used as a proxy for large subunit defects to identify a series of l10 mutants. these mapped to roughly four discrete regions of the protein. a det ...200818824477
kinetic and thermodynamic studies of peptidyltransferase in ribosomes from the extreme thermophile thermus thermophilus.throughout evolution, emerging organisms survived by adapting existing biochemical processes to new reaction conditions. simple protein enzymes balanced changes in structural stability with changes that permitted optimal catalysis by adjustments in both entropic and enthalpic contributions to the free energy of activation for the reaction. study of adaptive mechanisms by large multicomponent enzymes such as the ribosome has been largely unexplored. here we have determined the kinetic and thermod ...200818824514
identification and characterization of domains responsible for self-assembly and cell wall binding of the surface layer protein of lactobacillus brevis atcc 8287.lactobacillus brevis atcc 8287 is covered by a regular surface (s-) layer consisting of a 435 amino acid protein slpa. this protein is completely unrelated in sequence to the previously characterized s-layer proteins of lactobacillus acidophilus group.200818828902
the crystal structure of desulfovibrio vulgaris dissimilatory sulfite reductase bound to dsrc provides novel insights into the mechanism of sulfate respiration.sulfate reduction is one of the earliest types of energy metabolism used by ancestral organisms to sustain life. despite extensive studies, many questions remain about the way respiratory sulfate reduction is associated with energy conservation. a crucial enzyme in this process is the dissimilatory sulfite reductase (dsir), which contains a unique siroheme-[4fe4s] coupled cofactor. here, we report the structure of desulfoviridin from desulfovibrio vulgaris, in which the dsir dsrab (sulfite reduc ...200818829451
the mycoplasma pneumoniae mpn229 gene encodes a protein that selectively binds single-stranded dna and stimulates recombinase a-mediated dna strand exchange.mycoplasma pneumoniae has previously been characterized as a micro-organism that is genetically highly stable. in spite of this genetic stability, homologous dna recombination has been hypothesized to lie at the basis of antigenic variation of the major surface protein, p1, of m. pneumoniae. in order to identify the proteins that may be involved in homologous dna recombination in m. pneumoniae, we set out to characterize the mpn229 open reading frame (orf), which bears sequence similarity to the ...200818831760
ribosomal protein l3 functions as a 'rocker switch' to aid in coordinating of large subunit-associated functions in eukaryotes and archaea.although ribosomal rnas (rrnas) comprise the bulk of the ribosome and carry out its main functions, ribosomal proteins also appear to play important structural and functional roles. many ribosomal proteins contain long, nonglobular domains that extend deep into the rrna cores. in eukaryotes and archaea, ribosomal protein l3 contains two such extended domains tethered to a common globular hub, thus providing an excellent model to address basic questions relating to ribosomal protein structure/fun ...200818832371
structure of a sigma28-regulated nonflagellar virulence protein from campylobacter jejuni.campylobacter jejuni, a gram-negative motile bacterium, is a leading cause of human gastrointestinal infections. although the mechanism of c.jejuni-mediated enteritis appears to be multifactorial, flagella play complex roles in the virulence of this human pathogen. cj0977 is a recently identified virulence factor in c. jejuni and is expressed by a sigma(28) promoter that controls late genes in the flagellar regulon. a cj0977 mutant strain is fully motile but significantly reduced in the invasion ...200818835274
a unique combination of genetic systems for the synthesis of trehalose in rubrobacter xylanophilus: properties of a rare actinobacterial tret.trehalose is the primary organic solute in rubrobacter xylanophilus under all conditions tested, including those for optimal growth. we detected genes of four different pathways for trehalose synthesis in the genome of this organism, namely, the trehalose-6-phosphate synthase (tps)/trehalose-6-phosphate phosphatase (tpp), tres, trey/trez, and tret pathways. moreover, r. xylanophilus is the only known member of the phylum actinobacteria to harbor tret. the tps sequence is typically bacterial, but ...200818835983
the pretranslocation ribosome is targeted by gtp-bound ef-g in partially activated form.translocation of the trna x mrna complex through the bacterial ribosome is driven by the multidomain guanosine triphosphatase elongation factor g (ef-g). we have used isothermal titration calorimetry to characterize the binding of gdp and gtp to free ef-g at 4 degrees c, 20 degrees c, and 37 degrees c. the binding affinity of ef-g is higher to gdp than to gtp at 4 degrees c, but lower at 37 degrees c. the binding enthalpy and entropy change little with temperature in the case of gdp binding but ...200818836081
crystal structure of muts2 endonuclease domain and the mechanism of homologous recombination suppression.dna recombination events need to be strictly regulated, because an increase in the recombinational frequency causes unfavorable alteration of genetic information. recent studies revealed the existence of a novel anti-recombination enzyme, muts2. however, the mechanism by which muts2 inhibits homologous recombination has been unknown. previously, we found that thermus thermophilus muts2 (ttmuts2) harbors an endonuclease activity and that this activity is confined to the c-terminal domain, whose a ...200818838375
the higher level of organization of the oxidative phosphorylation system: mitochondrial supercomplexes.the organization of the oxidative phosphorylation (oxphos) system within the inner mitochondrial membrane appears to be far more complicated than previously thought. in particular, the individual protein complexes of the oxphos system (complexes i to v) were found to specifically interact forming defined supramolecular structures. blue-native polyacrylamide gel electrophoresis and single particle electron microscopy proved to be especially valuable in studying the so-called "respiratory supercom ...200818839290
conformationally gated metal uptake by apomanganese superoxide dismutase.metal uptake by apomanganese superoxide dismutase in vitro is a complex process exhibiting multiphase "gated" reaction kinetics and a striking sigmoidal temperature profile that has led to a model of conformationally gated metal binding, requiring conversion between "closed" and "open" forms. this work systematically explores the structural determinants of metal binding in both wild-type (wt) apoprotein and mutational variants as a test of mechanistic models. the ph dependence of metalation unde ...200818841998
rnomics and modomics in the halophilic archaea haloferax volcanii: identification of rna modification genes.naturally occurring rnas contain numerous enzymatically altered nucleosides. differences in rna populations (rnomics) and pattern of rna modifications (modomics) depends on the organism analyzed and are two of the criteria that distinguish the three kingdoms of life. if the genomic sequences of the rna molecules can be derived from whole genome sequence information, the modification profile cannot and requires or direct sequencing of the rnas or predictive methods base on the presence or absence ...200818844986
proton-dependent electron transfer from cua to heme a and altered epr spectra in mutants close to heme a of cytochrome oxidase.eukaryotic cytochrome c oxidase (cco) and homologous prokaryotic forms of rhodobacter and paraccocus differ in the epr spectrum of heme a. it was noted that a histidine ligand of heme a (h102) is hydrogen bonded to serine in rhodobacter (s44) and paraccocus ccos, in contrast to glycine in the bovine enzyme. mutation of s44 to glycine shifts the heme a epr signal from g(z) = 2.82 to 2.86, closer to bovine heme a at 3.03, without modifying other properties. mutation to aspartate, however, results ...200818847227
the high-resolution nmr structure of the early folding intermediate of the thermus thermophilus ribonuclease h.elucidation of the high-resolution structures of folding intermediates is a necessary but difficult step toward the ultimate understanding of the mechanism of protein folding. here, using hydrogen-exchange-directed protein engineering, we populated the folding intermediate of the thermus thermophilus ribonuclease h, which forms before the rate-limiting transition state, by removing the unfolded regions of the intermediate, including an alpha-helix and two beta-strands (51 folded residues). using ...200818848567
dna polymerases and aminoacyl-trna synthetases: shared mechanisms for ensuring the fidelity of gene expression.dna polymerases and aminoacyl-trna synthetases (arss) represent large enzyme families with critical roles in the transformation of genetic information from dna to rna to protein. dna polymerases carry out replication and collaborate in the repair of the genome, while arss provide aminoacylated trna precursors for protein synthesis. enzymes of both families face the common challenge of selecting their cognate small molecule substrates from a pool of chemically related molecules, achieving high le ...200818850722
the bet v 1 fold: an ancient, versatile scaffold for binding of large, hydrophobic ligands.the major birch pollen allergen, bet v 1, is a member of the ubiquitous pr-10 family of plant pathogenesis-related proteins. in recent years, a number of diverse plant proteins with low sequence similarity to bet v 1 was identified. in addition, determination of the bet v 1 structure revealed the existence of a large superfamily of structurally related proteins. in this study, we aimed to identify and classify all bet v 1-related structures from the protein data bank and all bet v 1-related sequ ...200818922149
conformational transition of sec machinery inferred from bacterial secye structures.over 30% of proteins are secreted across or integrated into membranes. their newly synthesized forms contain either cleavable signal sequences or non-cleavable membrane anchor sequences, which direct them to the evolutionarily conserved sec translocon (secyeg in prokaryotes and sec61, comprising alpha-, gamma- and beta-subunits, in eukaryotes). the translocon then functions as a protein-conducting channel. these processes of protein localization occur either at or after translation. in bacteria, ...200818923527
rare codons cluster.most amino acids are encoded by more than one codon. these synonymous codons are not used with equal frequency: in every organism, some codons are used more commonly, while others are more rare. though the encoded protein sequence is identical, selective pressures favor more common codons for enhanced translation speed and fidelity. however, rare codons persist, presumably due to neutral drift. here, we determine whether other, unknown factors, beyond neutral drift, affect the selection and/or d ...200818923675
the unique nature of mg2+ channels.considering the biological abundance and importance of mg2+, there is a surprising lack of information regarding the proteins that transport mg2+, the mechanisms by which they do so, and their physiological roles within the cell. the best characterized mg2+ channel to date is the bacterial protein cora, present in a wide range of bacterial species. the cora homolog mrs2 forms the mitochondrial mg2+ channel in all eukaryotes. physiologically, cora is involved in bacterial pathogenesis, and the mr ...200818927203
toward a chemical mechanism of proton pumping by the b-type cytochrome c oxidases: application of density functional theory to cytochrome ba3 of thermus thermophilus.a mechanism for proton pumping by the b-type cytochrome c oxidases is presented in which one proton is pumped in conjunction with the weakly exergonic, two-electron reduction of fe-bound o 2 to the fe-cu bridging peroxodianion and three protons are pumped in conjunction with the highly exergonic, two-electron reduction of fe(iii)- (-)o-o (-)-cu(ii) to form water and the active oxidized enzyme, fe(iii)- (-)oh,cu(ii). the scheme is based on the active-site structure of cytochrome ba 3 from thermus ...200818928258
purification, crystallization and preliminary x-ray diffraction analysis of the cbs-domain pair from the methanococcus jannaschii protein mj0100.cbs domains are small protein motifs consisting of a three-stranded beta-sheet and two alpha-helices that are present in proteins of all kingdoms of life and in proteins with completely different functions. several genetic diseases in humans have been associated with mutations in their sequence, which has made them promising targets for rational drug design. the c-terminal domain of the methanococcus jannaschii protein mj0100 includes a cbs-domain pair and has been overexpressed, purified and cr ...200818931440
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