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novel hybrid encodes both continuous and split trna genes?trnas are mostly transcribed from un-fragmented genes, but occasionally also from split genes, with separated 5' and 3' halves. a reanalysis of the existing data on staphylothermus marinus and staphylothermus hellenicus hints of a novel hybrid gene that encodes both an un-fragmented and a 5'-split-half together in one. the corresponding 3' complement-gene is located elsewhere on the genome. as un-fragmented, the hybrid gene transcribes to trna(lys)(ttt). but as 5'-half, it trans-splices with its ...201121294593
overexpression and characterization of an extremely thermostable maltogenic amylase, with an optimal temperature of 100 degrees c, from the hyperthermophilic archaeon staphylothermus marinus.a gene encoding a hyperthermostable maltogenic amylase of staphylothermus marinus (smma) was cloned and overexpressed in escherichia coli. smma consisted of 696 amino acids with a predicted molecular mass of 82.5 kda. the enzyme was active in acidic conditions (ph 3.5-5.0), with an optimal ph of 5.0, and was extremely thermostable, with a temperature optimum of 100 degrees c and a melting temperature of 109 degrees c, both of which extremely favored the starch conversion process. smma hydrolyzed ...201020385261
the use of coiled-coil proteins in drug delivery systems.the coiled-coil motif is found in approximately 10% of all protein sequences and is responsible for the oligomerization of proteins in a highly specific manner. coiled-coil proteins exhibit a large diversity of function (e.g. gene regulation, cell division, membrane fusion, drug extrusion) thus demonstrating the significance of oligomerization in biological systems. the classical coiled-coil domain comprises a series of consecutive heptad repeats in the protein sequence that are readily identifi ...200919835864
acidianus, sulfolobus and metallosphaera surface layers: structure, composition and gene expression.the cell walls of sulfolobales species consist of proteinaceous s-layers assembled from two polypeptides, slaa and slab. we isolated the large s-layer protein of acidianus ambivalens and both s-layer subunits of sulfolobus solfataricus and metallosphaera sedula, respectively. the slaab genes, lying adjacently in the chromosomes, are constitutively transcribed as bicistronic operons in a. ambivalens and s. solfataricus. a smaller slaa transcript appeared in northern hybridizations of a. ambivalen ...200919522740
the complete genome sequence of staphylothermus marinus reveals differences in sulfur metabolism among heterotrophic crenarchaeota.staphylothermus marinus is an anaerobic, sulfur-reducing peptide fermenter of the archaeal phylum crenarchaeota. it is the third heterotrophic, obligate sulfur reducing crenarchaeote to be sequenced and provides an opportunity for comparative analysis of the three genomes.200919341479
adpase activity of recombinantly expressed thermotolerant atpases may be caused by copurification of adenylate kinase of escherichia coli.except for apyrases, atpases generally target only the gamma-phosphate of a nucleotide. some non-apyrase atpases from thermophilic microorganisms are reported to hydrolyze adp as well as atp, which has been described as a novel property of the atpases from extreme thermophiles. here, we describe an apparent adp hydrolysis by highly purified preparations of the aaa+ atpase ntrc1 from an extremely thermophilic bacterium, aquifex aeolicus. this activity is actually a combination of the activities o ...200919143839
cellular polyamines of the acidophilic, thermophilic and thermoacidophilic archaebacteria, acidilobus, ferroplasma, pyrobaculum, pyrococcus, staphylothermus, thermococcus, thermodiscus and vulcanisaeta.cellular polyamines of newly isolated acidophilic, thermophilic and thermoacidophilic archaebacteria were investigated for the chemotaxonomic significance of polyamine distribution profiles. in addition to spermidine, spermine and agmatine, a quaternary branched penta-amine, n(4)-bis(aminopropyl)spermidine, was found in thermophilic thermococcus waiotapuensis, thermococcus aegaeus and pyrococcus glycovorans belonging to the order thermococcales. an acidophilic euryarchaeon, ferroplasma acidiphil ...200314673752
utilization of a right-handed coiled-coil protein from archaebacterium staphylothermus marinus as a carrier for cisplatin.the nano-sized right-handed coiled-coil (rhcc) protein, originating from the archaebacterium staphylothermus marinus, is stable at high salt concentrations, high temperatures, high pressures and extremes of ph. its crystal structure reveals four hydrophobic cavities which can incorporate heavy metals. nano-sized compounds have been used to carry cytotoxic drugs to tumours, avoiding delivery to healthy tissue, in part due to enhanced permeability in tumour blood vessels (enhanced permeability and ...200919331128
novel molecular architecture of the multimeric archaeal pep-synthase homologue (maps) from staphylothermus marinus.the phosphoenolpyruvate (pep)-synthases belong to the family of structurally and functionally related pep-utilizing enzymes. the only archaeal member of this family characterized thus far is the multimeric archaeal pep-synthase homologue from staphylothermus marinus (maps). this protein complex differs from the bacterial and eukaryotic representatives characterized to date in its homomultimeric, as opposed to dimeric or tetrameric, structure. we have probed the molecular architecture of maps usi ...199910388577
structural studies on the 2.25-mda homomultimeric phosphoenolpyruvate synthase from staphylothermus marinus.the phosphoenolpyruvate synthase of the hyperthermophilic archaeon staphylothermus marinus forms an unusually large homomultimeric complex of 93 kda subunits. electron image analysis of negatively stained and low-dose unstained preparations showed that the complex has a single, stable characteristic view and a well-preserved core with threefold rotational symmetry. the periphery of the assembly is composed of a nebulous, possibly flexible, component. mass measurements by scanning transmission el ...19968812985
hyperthermostable surface layer protein tetrabrachion from the archaebacterium staphylothermus marinus: evidence for the presence of a right-handed coiled coil derived from the primary structure.the scaffold of the surface layer covering the hyperthermophilic archaebacterium staphylothermus marinus is formed by an extended filiform glycoprotein complex, tetrabrachion, which is anchored in the cell membrane at one end of a 70 nm stalk and branches at the other end into four arms of 24 nm length. the arms from a canopy-like meshwork by end-to-end contacts, enclosing a "quasi-periplasmic space". the primary structure of the complex, obtained by an approach based entirely on the polymerase ...19968632466
tetrabrachion: a filamentous archaebacterial surface protein assembly of unusual structure and extreme stability.the surface (s-) layer of the hyperthermophilic archaebacterium staphylothermus marinus was isolated, dissected into separate domains by chemical and proteolytic methods, and analyzed by spectroscopic, electron microscopic and biochemical techniques. the s-layer is formed by a poorly ordered meshwork of branched, filiform morphological subunits resembling dandelion seed-heads. a morphological subunit (christened by us tetrabrachion) consists of a 70 nm long, almost perfectly straight stalk endin ...19957837271
primary structure of a multimeric protein, homologous to the pep-utilizing enzyme family and isolated from a hyperthermophilic archaebacterium.a large protein complex (approx. 2000 kda) was found in the cytosol of the hyperthermophilic archaebacterium staphylothermus marinus. the purified protein was shown to be a homomultimer of 93 kda subunits, the primary structure of which was determined by nucleotide sequence analysis. the protein belongs to the family of phosphoenolpyruvate-utilizing enzymes and represents the first member characterized in archaebacteria. its homomultimeric organisation differs from the typically dimeric structur ...19947805870
thermogladius shockii gen. nov., sp. nov., a hyperthermophilic crenarchaeote from yellowstone national park, usa.a hyperthermophilic heterotrophic archaeon (strain wb1) was isolated from a thermal pool in the washburn hot spring group of yellowstone national park, usa. wb1 is a coccus, 0.6-1.2 μm in diameter, with a tetragonal s-layer, vacuoles, and occasional stalk-like protrusions. growth is optimal at 84°c (range 64-93°c), ph 5-6 (range 3.5-8.5), and <1 g/l nacl (range 0-4.6 g/l nacl). tests of metabolic properties show the isolate to be a strict anaerobe that ferments complex organic substrates. phylog ...201020978744
ribosomal rna introns in archaea and evidence for rna conformational changes associated with splicing.the single 23s rrna gene of the archaeon staphylothermus marinus exhibits two introns which, at the rna level, are located in highly conserved regions of domains iv and v. the rna introns, which are 56 and 54 nucleotides long, respectively, can form single hairpin structures. in vivo, rna splicing occurs efficiently, whereas in vitro pre-rrna transcripts containing each intron were cleaved efficiently when incubated with archaeal cell extracts but were poorly ligated. the introns are cleaved by ...19911899138
structural features of archaebacterial cell envelopes.regularly arrayed surface (glyco) proteins--often referred to as s layers--are a common feature of the cell envelopes of almost all archaebacteria. we have selected some examples (halobacterium, sulfolobus, thermoproteus, pyrobaculum, staphylothermus), and we describe the structure of their surface layers as revealed primarily by electron crystallography. in spite of a considerable diversity in shapes and dimensions, some common structural features emerge from the comparison. the glycoprotein ar ...19921459988
complete genome sequence of staphylothermus marinus stetter and fiala 1986 type strain f1.staphylothermus marinus fiala and stetter 1986 belongs to the order desulfurococcales within the archaeal phylum crenarchaeota. s. marinus is a hyperthermophilic, sulfur-dependent, anaerobic heterotroph. strain f1 was isolated from geothermally heated sediments at vulcano, italy, but s. marinus has also been isolated from a hydrothermal vent on the east pacific rise. we report the complete genome of s. marinus strain f1, the type strain of the species. this is the fifth reported complete genome ...200921304655
thermococcus aegaeicus sp. nov. and staphylothermus hellenicus sp. nov., two novel hyperthermophilic archaea isolated from geothermally heated vents off palaeochori bay, milos, greece.two novel, hyperthermophilic, anaerobic, heterotrophic archaea were isolated from shallow hydrothermal vents off palaeochori bay, milos, greece. strain p5t (bk17s6-3-b2t) is an irregular coccus, with a single polar flagellum, growing optimally at 90 degrees c, ph 6 and 2% nacl. the dna g+c content was 45 mol%. due to its morphology, phylogenetic analyses based on 16s rrna gene sequencing, dna-dna hybridization experiments, physiological properties and nutritional features, this strain represents ...200011155985
three-dimensional cryoelectron microscopic reconstruction of the 2.25-mda homomultimeric phosphoenolpyruvate synthase from staphylothermus marinus.the phosphoenolpyruvate synthase (ec 2.7.9.2) of the hyperthermophilic archaeon staphylothermus marinus forms a 2.25 mda homomultimeric complex of 24 subunits. here, computational analysis of low-dose cryoelectron micrographs was used to ascertain that the major rotational symmetry axes were 2-fold, 3-fold, and 4-fold. these symmetry considerations were used to perform a three-dimensional reconstruction to a spatial resolution of 4 nm. this assembly has an octahedral architecture with a solvent ...19979425318
a hyperthermostable protease of the subtilisin family bound to the surface layer of the archaeon staphylothermus marinus.staphylothermus marinus, an archaeon isolated from a geothermally heated marine environment, is a peptide-fermenting, sulphur-dependent organism with an optimum growth temperature of 92 degrees c. it forms grapes of cells, which adhere to each other and to sulphur granules via their surface layer. this glycoprotein layer forms a canopy which is held at a distance of about 70 nm from the cell membrane by membrane-anchored stalks, thereby enclosing a 'quasi-periplasmic space'. two copies of a glob ...19968793300
complete genome sequence of staphylothermus hellenicus p8.staphylothermus hellenicus belongs to the order desulfurococcales within the archaeal phylum crenarchaeota. strain p8(t) is the type strain of the species and was isolated from a shallow hydrothermal vent system at palaeochori bay, milos, greece. it is a hyperthermophilic, anaerobic heterotroph. here we describe the features of this organism together with the complete genome sequence and annotation. the 1,580,347 bp genome with its 1,668 protein-coding and 48 rna genes was sequenced as part of a ...201122180806
association of a novel domain in the active site of archaic hyperthermophile maltogenic amylase from staphylothermus marinus.smma (staphylothermus marinus maltogenic amylase) is a novel extreme thermophile maltogenic amylase with an optimal temperature of 100°c, which hydrolyzes α-(1-4) glycosyl linkages in cyclodextrin and in inear malto-oligosaccharide. this enzyme has a long n-terminal extension that is conserved among archaic hyperthermophile amylases but is not found in other hydrolyzing enzymes from the gh13 family. the smma crystal structure revealed that the n-terminal extension forms an n' domain that is simi ...201222223643
crystal structure of a naturally occurring parallel right-handed coiled coil tetramer.the crystal structure of a polypeptide chain fragment from the surface layer protein tetrabrachion from staphylothermus marinus has been determined at 1.8 a resolution. as proposed on the basis of the presence of 11-residue repeats, the polypeptide chain fragment forms a parallel right-handed coiled coil structure. complementary hydrophobic interactions and complex networks of surface salt bridges result in an extremely thermostable tetrameric structure with remarkable properties. in marked cont ...200010966648
5'-methylbenzimidazolyl-cobamides are the corrinoids from some sulfate-reducing and sulfur-metabolizing bacteria.the sulfate-reducing bacteria desulfobacterium autotrophicum, desulfobulbus propionicus and archaeoglobus fulgidus (vc-16) and the sulfur-metabolizing archaebacteria desulfurolobus ambivalens and thermoplasma acidophilum were found to contain considerable amounts of corrinoids, that were isolated and crystallized in their co beta-cyano form. in three other sulfur-metabolizing archaebacteria, thermoproteus neutrophilus, pyrodictium occultum and staphylothermus marinus significant amounts of corri ...19883416881
angular reconstitution of the staphylothermus marinus phosphoenolpyruvate synthase.the processes of single particle electron crystallography and three-dimensional angular reconstitution are applied to digital cryoelectron images of a macromolecular complex, the staphylothermus marinus phosphoenolpyruvate synthase. in particular, the application of iqad (iterative quaternionic angular determination) is exemplified in the context of more canonical approaches.200010816265
cloning and expression of a dna ligase from the hyperthermophilic archaeon staphylothermus marinus and properties of the enzyme.the gene encoding staphylothermus marinus dna ligase (sma dna ligase) was cloned and sequenced. the gene contains an open reading frame consisting of 1836bp, which encodes for 611 amino acid residues. upon alignment of the entire amino acid sequence, sma dna ligase showed a high degree of sequence homology with the hyperthemophilic archaeal dna ligases, 67% identity with aeropyrum pernix k1, and 40% identity with both pyrococcus abyssi and thermococcus kodakarensis. an extremely high sequence id ...200717118474
minimal sulfur requirement for growth and sulfur-dependent metabolism of the hyperthermophilic archaeon staphylothermus marinus.staphylothermus marinus is an anaerobic hyperthermophilic archaeon that uses peptides as carbon and energy sources. elemental sulfur (s(o)) is obligately required for its growth and is reduced to h2s. the metabolic functions and mechanisms of s(o) reduction were explored by examining s(o)-dependent growth and activities of key enzymes present in this organism. all three forms of s(o) tested--sublimed s(o), colloidal s(o) and polysulfide--were used by s. marinus, and no other sulfur-containing co ...200315803665
quaternary organization of the staphylothermus marinus phosphoenolpyruvate synthase: angular reconstitution from cryoelectron micrographs with molecular modeling.digital electron images of frozen-hydrated preparations of the 2.25-mda staphylothermus marinus phosphoenolpyruvate synthase (ec 2.7.9.2) have been analyzed by single-particle classification and averaging and iterative quaternion-based angular reconstitution. contrast transfer function correction of micrographs obtained at different defocus values was used to improve the informational quality of the projection averages. three-dimensional reconstructions were obtained to roughly 3-nm spatial reso ...200011243891
discovery of permuted and recently split transfer rnas in archaea.abstract: background: as in eukaryotes, precursor transfer rnas in archaea often contain introns that are removed in trna maturation. two unrelated archaeal species display unique pre-trna processing complexity in the form of split trna genes, in which 2-3 segments of trnas are transcribed from different loci, then trans-spliced to form a mature trna. another rare type of pre-trna, found only in eukaryotic algae, is permuted, where the 3' half is encoded upstream of the 5' half, and must be proc ...201121489296
a generic system for the expression and purification of soluble and stable influenza neuraminidase.the influenza surface glycoprotein neuraminidase (na) is essential for the efficient spread of the virus. antiviral drugs such as tamiflu (oseltamivir) and relenza (zanamivir) that inhibit na enzyme activity have been shown to be effective in the treatment of influenza infections. the recent 'swine flu' pandemic and world-wide emergence of tamiflu-resistant seasonal human influenza a(h1n1) h(274)y have highlighted the need for the ongoing development of new anti-virals, efficient production of v ...201121326879
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