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191.
Lipolytic bacteria were isolated from two stations on Brewery Creek, an arm of the Ottawa River, during the winter of 1971-72. Total counts were approximately sevenfold higher at the more polluted downstream station, whereas lipolytic counts were about 100-fold higher. At this station, significantly more lipolytic bacteria grew on plates incubated at 20 C than at 4 C, suggesting that the population was comprised of both mesophiles and psychrophiles. However, at the upstream station, approximately the same number were obtained at both temperatures. A total of 434 isolates, mainly from the downstream station, were tentatively classified. The major groups were Pseudomonas, Acinetobacter-Moraxella, and Aeromonas. Though the total number of lipolytic bacteria was fairly constant throughout the winter, the relative abundance of the acinetobacters dropped from approximately 90% in November to less than 10% in March, and then increased. The aeromonads and pseudomonads showed the opposite trend. Most of the bacteria, though isolated at 4 C, also grew at 30 C. Lipolysis, however, was generally strongest at 20 C or below. 相似文献
192.
Blaise F. Frost Gary D. Small 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1984,782(2):170-176
The major AP endonuclease from Chlamydomonas reinhardi has been partially purified and characterized. The enzyme has a molecular weight of about 38 000 as measured by molecular sieving. There is an absolute requirement for a divalent cation, with magnesium being better than manganese. The activity is stimulated by dithiothreitol and Triton X-100. The activity is sensitive to ionic strength, as 50 mM NaCl or KCl results in 70% inhibition. The enzyme is specific for apurinic and apyrimidinic (AP) sites and does not cleave DNA that has been damaged by ultraviolet light, methyl methanesulfonate, osmium tetroxide or sodium bisulfite. There is no deficiency in the AP endonuclease activity in extracts prepared from two mutants of Chlamydomonas that are sensitive to both ultraviolet light and methyl methanesulfonate. There was no evidence for induction of AP endonuclease after exposure of the cells to methyl methanesulfonate. 相似文献
193.
Blaise M Fréchin M Oliéric V Charron C Sauter C Lorber B Roy H Kern D 《Journal of molecular biology》2011,412(3):437-452
Asparagine synthetase A (AsnA) catalyzes asparagine synthesis using aspartate, ATP, and ammonia as substrates. Asparagine is formed in two steps: the β-carboxylate group of aspartate is first activated by ATP to form an aminoacyl-AMP before its amidation by a nucleophilic attack with an ammonium ion. Interestingly, this mechanism of amino acid activation resembles that used by aminoacyl-tRNA synthetases, which first activate the α-carboxylate group of the amino acid to form also an aminoacyl-AMP before they transfer the activated amino acid onto the cognate tRNA. In a previous investigation, we have shown that the open reading frame of Pyrococcus abyssi annotated as asparaginyl-tRNA synthetase (AsnRS) 2 is, in fact, an archaeal asparagine synthetase A (AS-AR) that evolved from an ancestral aspartyl-tRNA synthetase (AspRS). We present here the crystal structure of this AS-AR. The fold of this protein is similar to that of bacterial AsnA and resembles the catalytic cores of AspRS and AsnRS. The high-resolution structures of AS-AR associated with its substrates and end-products help to understand the reaction mechanism of asparagine formation and release. A comparison of the catalytic core of AS-AR with those of archaeal AspRS and AsnRS and with that of bacterial AsnA reveals a strong conservation. This study uncovers how the active site of the ancestral AspRS rearranged throughout evolution to transform an enzyme activating the α-carboxylate group into an enzyme that is able to activate the β-carboxylate group of aspartate, which can react with ammonia instead of tRNA. 相似文献
194.
Industrial strains of a polyploid, distiller’s Saccharomyces cerevisiae that produces glucoamylase and α-amylase was used for the direct fermentation of raw starch to ethanol. Strains contained
either Aspergillus awamori glucoamylase gene (GA1), Debaryomyces occidentalis glucoamylase gene (GAM1) or D. occidentalis α-amylase gene (AMY), singly or in combination, integrated into their chromosomes. The strain expressing both GA1 and AMY generated 10.3% (v/v) ethanol (80.9 g l−1) from 20% (w/v) raw corn starch after 6 days of fermentation, and decreased the raw starch content to 21% of the initial
concentration. 相似文献
195.
Daniel Raimunda Manuel González-Guerrero Blaise W. LeeberIII José M. Argüello 《Biometals》2011,24(3):467-475
Cu+-ATPases play a key role in bacterial Cu+ homeostasis by participating in Cu+ detoxification and cuproprotein assembly. Characterization of Archaeoglobus fulgidus CopA, a model protein within the subfamily of P1B-1 type ATPases, has provided structural and mechanistic details on this group of transporters. Atomic resolution structures
of cytoplasmic regulatory metal binding domains (MBDs) and catalytic actuator, phosphorylation, and nucleotide binding domains
are available. These, in combination with whole protein structures resulting from cryo-electron microscopy analyses, have
enabled the initial modeling of these transporters. Invariant residues in helixes 6, 7 and 8 form two transmembrane metal
binding sites (TM-MBSs). These bind Cu+ with high affinity in a trigonal planar geometry. The cytoplasmic Cu+ chaperone CopZ transfers the metal directly to the TM-MBSs; however, loading both of the TM-MBSs requires binding of nucleotides
to the enzyme. In agreement with the classical transport mechanism of P-type ATPases, occupancy of both transmembrane sites
by cytoplasmic Cu+ is a requirement for enzyme phosphorylation and subsequent transport into the periplasmic or extracellular milieus. Recent
transport studies have shown that all Cu+-ATPases drive cytoplasmic Cu+ efflux, albeit with quite different transport rates in tune with their various physiological roles. Archetypical Cu+-efflux pumps responsible for Cu+ tolerance, like the Escherichia coli CopA, have turnover rates ten times higher than those involved in cuproprotein assembly (or alternative functions). This
explains the incapability of the latter group to significantly contribute to the metal efflux required for survival in high
copper environments. 相似文献
196.
Blüggel M Spertini F Lutter P Wassenberg J Audran R Corthésy B Müllner S Blum S Wattenberg A Mercenier A Affolter M Kussmann M 《Journal of proteome research》2011,10(4):1558-1570
Allergy is an immunological disorder of the upper airways, lung, skin, and the gut with a growing prevalence over the last decades in Western countries. Atopy, the genetic predisposition for allergy, is strongly dependent on familial inheritance and environmental factors. These observations call for predictive markers of progression from atopy to allergy, a prerequisite to any active intervention in neonates and children (prophylactic interventions/primary prevention) or in adults (immunomodulatory interventions/secondary prevention). In an attempt to identify early biomarkers of the "atopic march" using minimally invasive sampling, CD4+ T cells from 20 adult volunteers (10 healthy and 10 with respiratory allergies) were isolated and quantitatively analyzed and their proteomes were compared in and out of pollen season (± antigen exposure). The proteome study based on high-resolution 2D gel electrophoresis revealed three candidate protein markers that distinguish the CD4+ T cell proteomes of normal from allergic individuals when sampled out of pollen season, namely Talin 1, Nipsnap homologue 3A, and Glutamate-cysteine ligase regulatory protein. Three proteins were found differentially expressed between the CD4+ T cell proteomes of normal and allergic subjects when sampled during pollen season: carbonyl reductase, glutathione S-transferase ω 1, and 2,4-dienoyl-CoA reductase. The results were partly validated by Western blotting. 相似文献
197.
Amélie Vromman Nesrine Trabelsi Clotilde Rouxel Gilbert Béréziat Isabelle Limon Régis Blaise 《Aging cell》2013,12(3):358-369
Several studies have shown that the accumulation of β‐amyloid peptides in the brain parenchyma or vessel wall generates an inflammatory environment. Some even suggest that there is a cause‐and‐effect relationship between inflammation and the development of Alzheimer's disease and/or cerebral amyloid angiopathy (CAA). Here, we studied the ability of wild‐type Aβ1‐40‐peptide (the main amyloid peptide that accumulates in the vessel wall in sporadic forms of CAA) to modulate the phenotypic transition of vascular smooth muscle cells (VSMCs) toward an inflammatory/de‐differentiated state. We found that Aβ1‐40‐peptide alone neither induces an inflammatory response, nor decreases the expression of contractile markers; however, the inflammatory response of VSMCs exposed to Aβ1‐40‐peptide prior to the addition of the pro‐inflammatory cytokine IL‐1β is greatly intensified compared with IL‐1β‐treated VSMCs previously un‐exposed to Aβ1‐40‐peptide. Similar conclusions could be drawn when tracking the decline of contractile markers. Furthermore, we found that the mechanism of this potentiation highly depends on an Aβ1‐40 preactivation of the PI3Kinase and possibly NFκB pathway; indeed, blocking the activation of these pathways during Aβ1‐40‐peptide treatment completely suppressed the observed potentiation. Finally, strengthening the possible in vivo relevance of our findings, we evidenced that endothelial cells exposed to Aβ1‐40‐peptide generate an inflammatory context and have similar effects than the ones described with IL‐1β. These results reinforce the idea that intraparietal amyloid deposits triggering adhesion molecules in endothelial cells, contribute to the transition of VSMCs to an inflammatory/de‐differentiated phenotype. Therefore, we suggest that acute inflammatory episodes may increase vascular alterations and contribute to the ontogenesis of CAA. 相似文献
198.
Sutapa Ray Victor Banerjee Mickael Blaise Baisakhi Banerjee Kali Pada Das Daniel Kern Rajat Banerjee 《The protein journal》2014,33(2):143-149
Glutamyl-queuosine-tRNAAsp synthetase (Glu-Q-RS) and glutamyl-tRNA synthetase (GluRS), differ widely by their function although they share close structural resemblance within their catalytic core of GluRS. In particular both Escherichia coli GluRS and Glu-Q-RS contain a single zinc-binding site in their putative tRNA acceptor stem-binding domain. It has been shown that the zinc is crucial for correct positioning of the tRNAGlu acceptor-end in the active site of E. coli GluRS. To address the role of zinc ion in Glu-Q-RS, the C101S/C103S Glu-Q-RS variant is constructed. Energy dispersive X-ray fluorescence show that the zinc ion still remained coordinated but the variant became structurally labile and acquired aggregation capacity. The extent of aggregation of the protein is significantly decreased in presence of the small substrates and more particularly by adenosine triphosphate. Addition of zinc increased significantly the solubility of the variant. The aminoacylation assay reveals a decrease in activity of the variant even after addition of zinc as compared to the wild-type, although the secondary structure of the protein is not altered as shown by the Fourier transform infrared spectroscopy study. 相似文献
199.
Enhanced intracellular survival (Eis) proteins belonging to the superfamily of the GCN5‐related N‐acetyltransferases play important functions in mycobacterial pathogenesis. In Mycobacterium tuberculosis, Eis enhances the intracellular survival of the bacilli in macrophages by modulating the host immune response and is capable to chemically modify and inactivate aminoglycosides. In nontuberculous mycobacteria (NTM), Eis shares similar functions. However, Mycobacterium abscessus, a multidrug resistant NTM, possesses two functionally distinct Eis homologues, Eis1Mab and Eis2Mab. While Eis2Mab participates in virulence and aminoglycosides resistance, this is not the case for Eis1Mab, whose exact biological function remains to be determined. Herein, we show that overexpression of Eis1Mab in M. abscessus fails to induce resistance to aminoglycosides. To clarify why Eis1Mab is unable to modify this class of antibiotics, we solved its crystal structure bound to its cofactor, acetyl‐CoA. The structure revealed that Eis1Mab has a typical homohexameric Eis‐like organization. The structural analysis supported by biochemical approaches demonstrated that while Eis1Mab can acetylate small substrates, its active site is too narrow to accommodate aminoglycosides. Comparison with other Eis structures showed that an extended loop between strands 9 and 10 is blocking the access of large substrates to the active site and movement of helices 4 and 5 reduces the volume of the substrate‐binding pocket to these compounds in Eis1Mab. Overall, this study underscores the molecular determinants explaining functional differences between Eis1Mab and Eis2Mab, especially those inherent to their capacity to modify aminoglycosides. 相似文献
200.