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Quevillon-Cheruel S Leulliot N Meyer P Graille M Bremang M Blondeau K Sorel I Poupon A Janin J van Tilbeurgh H 《The Journal of biological chemistry》2004,279(1):619-625
Chorismate synthase (EC 4.2.3.5), the seventh enzyme in the shikimate pathway, catalyzes the transformation of 5-enolpyruvylshikimate 3-phosphate (EPSP) to chorismate, which is the last common precursor in the biosynthesis of numerous aromatic compounds in bacteria, fungi, and plants. The chorismate synthase reaction involves a 1,4-trans-elimination of phosphoric acid from EPSP and has an absolute requirement for reduced FMN as a cofactor. We have determined the three-dimensional x-ray structure of the yeast chorismate synthase from selenomethionine-labeled crystals at 2.2-A resolution. The structure shows a novel betaalphabetaalpha fold consisting of an alternate tight packing of two alpha-helical and two beta-sheet layers, showing no resemblance to any documented protein structure. The molecule is arranged as a tight tetramer with D2 symmetry, in accordance with its quaternary structure in solution. Electron density is missing for 23% of the amino acids, spread over sequence regions that in the three-dimensional structure converge on the surface of the protein. Many totally conserved residues are contained within these regions, and they probably form a structured but mobile domain that closes over a cleft upon substrate binding and catalysis. This hypothesis is supported by previously published spectroscopic measurements implying that the enzyme undergoes considerable structural changes upon binding of both FMN and EPSP. 相似文献
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Yolanda M. Drozdowicz Yu-Ping Lu Vijay Patel Sorel Fitz-Gibbon Jeffrey H. Miller Philip A. Rea 《FEBS letters》1999,460(3)
Vacuolar-type H+-translocating pyrophosphatases (V-PPases) have been considered to be restricted to plants, a few species of phototrophic proteobacteria and protists. Here, we describe PVP, a thermostable, sequence-divergent V-PPase from the facultatively aerobic hyperthermophilic archaeon Pyrobaculum aerophilum. PVP shares only 38% sequence identity with both the prototypical V-PPase from Arabidopsis thaliana and the H+-PPi synthase from Rhodospirillum rubrum, yet possesses most of the structural features characteristic of V-PPases. Heterologous expression of PVP in Saccharomyces cerevisiae yields a Mr 64 000 membrane polypeptide that specifically catalyzes Mg2+-dependent PPi hydrolysis. The existence of PVP implies that PPi-energized H+-translocation is phylogenetically more deeply rooted than previously thought. 相似文献
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Vacuolar-type H(+)-translocating pyrophosphatases (V-PPases) have been considered to be restricted to plants, a few species of phototrophic proteobacteria and protists. Here, we describe PVP, a thermostable, sequence-divergent V-PPase from the facultatively aerobic hyperthermophilic archaeon Pyrobaculum aerophilum. PVP shares only 38% sequence identity with both the prototypical V-PPase from Arabidopsis thaliana and the H(+)-PPi synthase from Rhodospirillum rubrum, yet possesses most of the structural features characteristic of V-PPases. Heterologous expression of PVP in Saccharomyces cerevisiae yields a M(r) 64? omitted?000 membrane polypeptide that specifically catalyzes Mg(2+)-dependent PPi hydrolysis. The existence of PVP implies that PPi-energized H(+)-translocation is phylogenetically more deeply rooted than previously thought. 相似文献
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A phylogenetic 'tree of life' has been constructed based on the observed presence and absence of families of protein-encoding genes observed in 11 complete genomes of free-living microorganisms. Past attempts to reconstruct the evolutionary relation-ships of microorganisms have been limited to sets of genes rather than complete genomes. Despite apparent rampant lateral gene transfer among microorganisms, these results indicate a single robust underlying evolutionary history for these organisms. Broadly, the tree produced is very similar to the small subunit rRNA tree although several additional phylogenetic relationships appear to be resolved, including the relationship of Archaeoglobus to the methanogens studied. This result is in contrast to notions that a robust phylogenetic reconstruction of microorganisms is impossible due to their genomes being composed of an incomprehensible amalgam of genes with complicated histories and suggests that this style of genome-wide phylogenetic analysis could become an important method for studying the ancient diversification of life on Earth. Analyses using informational and operational subsets of the genes showed that this 'tree of life' is not dependent on the phylogenetically more consistent informational genes. 相似文献
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Matthew E. Rhodes Sorel T. Fitz‐Gibbon Aharon Oren Christopher H. House 《Environmental microbiology》2010,12(9):2613-2623
The increase of the acidic nature of proteins as an adaptation to hypersalinity has been well documented within halophile isolates. Here we explore the effect of salinity on amino acid preference on an environmental scale. Via pyrosequencing, we have obtained two distinct metagenomic data sets from the Dead Sea, one from a 1992 archaeal bloom and one from the modern Dead Sea. Our data, along with metagenomes from environments representing a range of salinities, show a strong linear correlation (R2 = 0.97) between the salinity of an environment and the ratio of acidic to basic amino acids encoded by its inhabitants. Using the amino acid composition of putative protein‐encoding reads and the results of 16S rRNA amplicon sequencing, we differentiate recovered sequences representing microorganisms indigenous to the Dead Sea from lateral gene transfer events and foreign DNA. Our methods demonstrate lateral gene transfer events between a halophilic archaeon and relatives of the thermophilic bacterial genus Thermotoga and suggest the presence of indigenous Dead Sea representatives from 10 traditionally non‐hyperhalophilic bacterial lineages. The work suggests the possibility that amino acid bias of hypersaline environments might be preservable in fossil DNA or fossil amino acids, serving as a proxy for the salinity of an ancient environment. Finally, both the amino acid profile of the 2007 Dead Sea metagenome and the V9 amplicon library support the conclusion that the dominant microorganism inhabiting the Dead Sea is most closely related to a thus far uncultured relative of an alkaliphilic haloarchaeon. 相似文献
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Elkins JS Whitmer RA Sidney S Sorel M Yaffe K Johnston SC 《Obesity (Silver Spring, Md.)》2006,14(8):1472-1478
Objective: Obesity is a growing problem among middle‐aged individuals. We investigated whether obesity in middle‐aged individuals influences the need for future nursing home care and whether the risk of nursing home admission associated with obesity is greater in whites than in blacks. Research Methods and Procedures: The study population (N = 8804) consisted of long‐term members of the Kaiser Permanente Medical Care Plan ages 75 to 85 years in 1995 who had completed a standardized, multiphasic health checkup while in their 50s. The multiphasic health checkup examinations were performed as part of routine medical care between the years 1964 and 1973 and included standardized measurements of BMI. We used health plan records to assess incident nursing home admissions from 1995 to 2002. The risk of nursing home admission associated with standard categories of midlife BMI was estimated using Cox proportional hazard analysis. Results: During an average follow‐up of 5.1 years, the nursing home admission rate was 6.8 per 100 person‐years of observation. After adjustment for comorbidities, midlife obesity predicted incident nursing home admission ~25 years later [hazard ratio (HR), 1.30; 95% confidence interval (CI), 1.15 to 1.46; p < 0.001]. Overweight BMI at midlife was not associated with future nursing home admission (HR, 1.05; 95% CI, 0.97 to 1.14; p = 0.23). The risk of nursing home admission associated with midlife obesity was higher in whites (HR, 1.34; 95% CI, 1.17 to 1.54; p < 0.001) than in blacks (HR 1.15; 95% CI, 0.87 to 1.52; p = 0.32), but the difference between races was not significant (p for interaction = 0.65). Discussion: Obesity among middle‐aged individuals is associated with an increased risk of nursing home admission in late life and may be an important target for reducing the future societal burden of nursing home care. 相似文献
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Leulliot N Quevillon-Cheruel S Sorel I Li de La Sierra-Gallay I Collinet B Graille M Blondeau K Bettache N Poupon A Janin J van Tilbeurgh H 《The Journal of biological chemistry》2004,279(9):8351-8358
The important role of the serine/threonine protein phosphatase 2A (PP2A) in various cellular processes requires a precise and dynamic regulation of PP2A activity, localization, and substrate specificity. The regulation of the function of PP2A involves the reversible methylation of the COOH group of the C-terminal leucine of the catalytic subunit, which, in turn, controls the enzyme's heteromultimeric composition and confers different protein recognition and substrate specificity. We have determined the structure of PPM1, the yeast methyltransferase responsible for methylation of PP2A. The structure of PPM1 reveals a common S-adenosyl-l-methionine-dependent methyltransferase fold, with several insertions conferring the specific function and substrate recognition. The complexes with the S-adenosyl-l-methionine methyl donor and the S-adenosyl-l-homocysteine product and inhibitor unambiguously revealed the co-substrate binding site and provided a convincing hypothesis for the PP2A C-terminal peptide binding site. The structure of PPM1 in a second crystal form provides clues to the dynamic nature of the PPM1/PP2A interaction. 相似文献
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Li de La Sierra-Gallay I Collinet B Graille M Quevillon-Cheruel S Liger D Minard P Blondeau K Henckes G Aufrère R Leulliot N Zhou CZ Sorel I Ferrer JL Poupon A Janin J van Tilbeurgh H 《Proteins》2004,54(4):776-783
The protein product of the YGR205w gene of Saccharomyces cerevisiae was targeted as part of our yeast structural genomics project. YGR205w codes for a small (290 amino acids) protein with unknown structure and function. The only recognizable sequence feature is the presence of a Walker A motif (P loop) indicating a possible nucleotide binding/converting function. We determined the three-dimensional crystal structure of Se-methionine substituted protein using multiple anomalous diffraction. The structure revealed a well known mononucleotide fold and strong resemblance to the structure of small metabolite phosphorylating enzymes such as pantothenate and phosphoribulo kinase. Biochemical experiments show that YGR205w binds specifically ATP and, less tightly, ADP. The structure also revealed the presence of two bound sulphate ions, occupying opposite niches in a canyon that corresponds to the active site of the protein. One sulphate is bound to the P-loop in a position that corresponds to the position of beta-phosphate in mononucleotide protein ATP complex, suggesting the protein is indeed a kinase. The nature of the phosphate accepting substrate remains to be determined. 相似文献