directed mutagenesis of nicotinic receptors to investigate receptor function
Tài liệu Báo cáo khoa học: Site-directed mutagenesis of a loop at the active site of E1 (a2b2) of the pyruvate dehydrogenase complex A possible common sequence motif docx
... Temperature-dependence of catalytic activity The dependence of the catalytic activity of E1 on temperature was investigated using the DCPIP assay over a temperature range of 15 °C to 85 °C Assay mixtures ... activity of E1 mutants was investigated using three different assays: the DCPIP assay, which measures E1 activity by monitoring the reduction of DCPIP as an artificial electron acceptor instead of lipoamide; ... activity of the E1aD276A mutant increased steadily to reach twice that of wild-type E1 at 65 °C, after which the enzyme was inactivated Binding of E1 to the peripheral subunit-binding domain of E2...
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Báo cáo khoa học: Site-directed mutagenesis of selected residues at the active site of aryl-alcohol oxidase, an H2O2-producing ligninolytic enzyme pot
... 50% of them clustered together in front of the rectus (re)-face of the isoalloxazine ring of the FAD cofactor This substrate location is shown in Fig 2A, which includes the 10 molecules of veratryl ... formation) by an active site base contributing to the transfer of a hydride from the substrate a-carbon to the flavin cofactor [40–46] Site -directed mutagenesis suggested that the conserved histidine ... Site -directed mutagenesis of aryl-alcohol oxidase P Ferreira et al entry 1QJN) [19] Polar hydrogen atoms were added to the molecular model according to the valence and isoelectric point of each...
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Báo cáo khoa học: Characterization of HbpR binding by site-directed mutagenesis of its DNA-binding site and by deletion of the effector domain pot
... was impaired in binding of the R13 region Both of these patterns contributed to the loss of binding contacts of mutant 72 Construction of a DA HbpR derivative In order to study whether the A-domain ... operator DNA in vitro without effector recognition or ATP binding The HbpR protein even seems to be able to form an oligomeric complex capable of protecting the operator DNA in the absence of its ... representation of the relative densities of the free DNA measured at increasing concentrations of HbpR The percentage of free DNA was calculated from densitometric measurements of the radiolabeled...
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Báo cáo khoa học: Probing the catalytic potential of the hamster arylamine N-acetyltransferase 2 catalytic triad by site-directed mutagenesis of the proximal conserved residue, Tyr190 pot
... Site -directed mutagenesis, protein expression and purication Presteady-state kinetic parameters for the acetylation of NAT Site -directed mutagenesis of the hamster NAT2 Tyr190 to Phe (Y190F), to ... in losses of activity of 88% and 95%, respectively, relative to wild-type enzyme Presteady-state kinetics of NAT acetylation The rate of acetylation of NATs was determined with a stopped-ow apparatus ... assigned to a probable conformational change [26] In the case of Y190F, the value of log (k2 Kmacetyl) rose as a function of pH until a plateau was reached above pH 7.5 The data were t into a one-pKa...
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Báo cáo khoa học: Probing the mechanism of the bifunctional enzyme ketol-acid reductoisomerase by site-directed mutagenesis of the active site pot
... activity of E coli KARI towards 2-acetolactate and various 2-ketoacids The value of kcat (s)1) is similar to the Vm (UÆmg)1) because the subunit molecular mass of 59.5 (kDa) is almost equal to the ... the number of seconds in one minute Substrate Vm(UÆmg)1) kcat(s)1) 2-acetolactate 2-Ketopantoate 2-Ketoisovalerate 2-Ketovalerate 2-Ketobutyrate Pyruvate 3-hydroxypyruvate 3-Hydroxy-2-ketobutyrate ... near irreversibility of the conversion of the 2-ketoacid to TS2 However the reaction is inefficient due to slow conversion of TS2 to TS1 600 HMKB and racemic 3-hydroxy-2-ketobutyrate were prepared...
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Aerobic Uptake of Cholesterol by Ergosterol Auxotrophic Strains in Candidaglabrata & Random and Site-Directed Mutagenesis of ERG25 in Saccharomyces cerevisiae
... scheme of random mutagenesis “insert” creation 27 2.2 Diagrammatic scheme of random mutagenesis “vector” creation .29 3.1 GC profile of wild type Candida glabrata 39 3.2 GC profile of ... 40 3.3 GC profile of ERG7 in Candida glabrata 41 3.4 GC profile of ERG11 in Candida glabrata 42 3.5 GC profile of ERG25 in Candida glabrata 43 3.6 GC profile of ERG27 in ... addition of 60 µg/ml of ampicillin for selection LB media consisted of 10 g of pancreatic digest of casein, g of yeast extract, and 10 g/L of sodium chloride (46) dissolved in milli-Q water and autoclaved...
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Báo cáo Y học: Puri®cation and characterization of the human adenosine A2a receptor functionally expressed in Escherichia coli doc
... in receptor concentrations of 10±20 nmol per L of culture, or 1000±2000 receptors per cell This corresponds to 17±34 pmol receptor per mg of membrane protein Constructs coding for full-length receptors ... level of a given receptor is dif®cult to predict, we performed an initial screen, expressing the cDNAs of four different human GPCRs (somatostatin receptors S2 and S4, and adenosine receptors ... were investigated In E coli, all four receptors were N-terminally fused to MBP, whereas in the yeast Pichia pastoris they were N-terminally fused to the a-factor prepropeptide (S2 and A1 receptors...
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Báo cáo khoa học: Binding areas of urokinase-type plasminogen activator– plasminogen activator inhibitor-1 complex for endocytosis receptors of the low-density lipoprotein receptor family, determined by site-directed mutagenesis doc
... activator-plasminogen activator inhibitor-1 complex to the endocytosis receptors alpha2-macroglobulin receptor low-density lipoprotein receptor- related protein and very-low-density lipoprotein receptor ... for these receptors, the uPAPAI-1 complex binds to LRP-1A with an afnity much higher than that of the free components Thus, the Kd value for binding of the uPAPAI-1 complex to these receptors is ... identied 11 of importance for binding of the uPAPAI-1 complex to one, two, or all three receptors (Fig 2) However, the involvement of Lys124 in receptor binding could not be conrmed Screening of effects...
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báo cáo hóa học: " Evaluation of 18F-nifene binding to a4b2 nicotinic receptors in the rat brain using microPET imaging" ppt
... order to image the a4b2 nicotinic receptors This is useful in the assessment of nicotinic receptors in neurological diseases MicroPET studies in rats validated the faster binding profile of 18 ... Committee of the University of California, Irvine O N H 18 Figure Chemical structure of F 18 N F-nifene The focus in this work is on in vivo evaluation of 18Fnifene binding to a4b2 nicotinic receptors ... F-nifene occupancy with respect to nicotine At low doses of nicotine, 0.02 mg/kg, > 40% of receptors were occupied while at high doses (0.5 mg/kg) > 80% receptors were occupied with nicotine...
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Indoor tests to investigate the effect of brine depth on the performance of solar still
... The objective of this work is to investigate the effect of brine depth on the productivity and efficiency of solar still under laboratory conditions Table shows a summary of some of the studies ... plate of galvanized iron to prevent inflation of the mica as it becomes heated during the operation The three heaters were connected in series to supply a total current of 4.5 A, a voltage of 220 ... V and a total power flux of approximately 1000 W/m² to simulate the solar radiation intensity A variac was used to regulate the input voltage to the heaters Figure shows a photograph of the constructed...
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A computational study to investigate the effects of insulation and EGR in a diesel engine
... fuel consumption (BSFC) of the IDI engine to increase and the total engine efficiency to decrease compared to that of a DI diesel engine Because of these disadvantages of the IDI diesel engines, ... side of main chamber This leads to the formation of two large eddies each occupying a half of the main chamber and staying centered with respect to the two half of the bowl(don’t shown contours) ... weaker due to opposing flow in glow plug channel (don’t shown contours) Therefore, at the end of injection period at 380°CA the flame reaches near to injector location without distortion Also...
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This questionnaire is designed to investigate teachers’ attitude towards the applicability of process approach in teaching writi
... researcher of this thesis unselected them to avoid causing too much inconvenience to those students There are 10 teachers of English at the school and all of them were selected as the subjects of the ... stated in the research, the researcher tried to look into “some pending matters of theory and practice” with the hope to contribute to the improvement of ESP teaching- learning in general and business ... a letter of complaint Writing a letter of confirmation Writing a formal letter of giving direction In sum, the result of the first section of the questionnaire has revealed that most of the teachers...
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Tài liệu Báo cáo khoa học: EGF receptor in relation to tumor development: molecular basis of responsiveness of cancer cells to EGFR-targeting tyrosine kinase inhibitors docx
... adenocarcinomas induced in mice by mutant EGF receptors found in human lung cancers respond to a tyrosine kinase inhibitor or to down-regulation of the receptors Genes Dev 20, 1496–1510 Ji H, Li D, ... growth factor receptors have been identified as major mechanisms of acquired resistance to TKIs [78] However, it is still important to identify additional mechanisms of resistance and to overcome ... induces apoptosis in a Bax-dependent manner Inhibitor of apoptosis protein family Induction of MOMP following apoptotic stimuli results in the release of cytochrome c and subsequent activation of caspase-9...
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Tài liệu Báo cáo khoa học: Thermodynamic characterization of interleukin-8 monomer binding to CXCR1 receptor N-terminal domain ppt
... protein-coupled receptors I Diversity of receptor ligand interactions J Biol Chem 273, 17299–17302 Leong SR, Kabakoff RC & Hebert CA (1994) Complete mutagenesis of the extracellular domain of interleukin-8 ... are difficult to obtain because of their membrane-embedded state IL-8 and all other chemokine receptors share some unique properties compared with other members of GPCR class A receptors Chemokines ... that some of the acidic residues, in addition to the apolar residues, are involved in binding to neutrophil receptors [20] Although the results from the mutational studies of the receptor N-domain...
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Tài liệu Báo cáo khoa học: Computational approaches to understand a-conotoxin interactions at neuronal nicotinic receptors doc
... Changeux, J.P (2000) Nicotinic receptors at the amino acid level Annu Rev Pharmacol Toxicol 40, 431–458 Karlin, A (2002) Emerging structure of the nicotinic acetylcholine receptors Nat Rev Neurosci ... with nicotinic acetylcholine receptor maturation Biochemistry 41, 12329–12335 26 Schapira, M., Abagyan, R & Totrov, M (2002) Structural model of nicotinic acetylcholine receptor isotypes bound to ... protein reveals the ligand-binding domain of nicotinic receptors Nature 411, 269–276 Dwoskin, L.P & Crooks, P.A (2001) Competitive neuronal nicotinic receptor antagonists: a new direction for drug...
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Tài liệu Báo cáo khoa học: nsights into the reaction mechanism of glycosyl hydrolase family 49 Site-directed mutagenesis and substrate preference of isopullulanase doc
... respectively) were forced to be placed based on the position of isomaltose bound in the subsites +1 and +2 of Dex49A, although IPU does not produce isomaltose IPU was predicted to consist of two domains, ... isomaltose While the study of site -directed mutagenesis described above was carried out, a crystal structure of Penicillium minioluteum dextranase, Dex49A, complexed with a product, isomaltose, ... composed of residues 25–540 The overall structure of the model of IPU and the mutated residues in this study are shown in Fig To elucidate the mechanism of the substrate recognition of IPU, two...
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Tài liệu Báo cáo khoa học: Identification of critical active-site residues in angiotensin-converting enzyme-2 (ACE2) by site-directed mutagenesis docx
... the substrate ⁄ inhibitor This study provides the first investigation of the importance of key active-site residues of ACE2 through site -directed mutagenesis, with the aim of providing practical ... [21], and is hence proposed to be involved in binding of the C-terminus of peptide substrates (Fig 1A) To test this hypothesis, we used site -directed mutagenesis to replace the arginine with ... Schematic view of the active site of ACE2 and tACE Binding interactions of the inhibitor (A) MLN-4760 at the active site of ACE2 and (B) lisinopril at the active site of tACE Hydrogen bonds to the ligand...
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Tài liệu Báo cáo khoa học: Insights into the interaction of human arginase II with substrate and manganese ions by site-directed mutagenesis and kinetic studies Alteration of substrate specificity by replacement of Asn149 with Asp docx
... binding of the a-carboxylate and a-amino groups of BEC were also ascribed to the larger volume of the active site cleft of arginase II, which allows more water-mediated enzyme–inhibitor interactions ... affinity of metal binding to arginase II To further examine the effects of the His120fiAsn and His145fiAsn mutations on the interaction of the enzyme with manganese ions, maximally activated species of ... (Fig 3) The possibility of interference from the endogenous agmatinase of the bacterial vector was excluded by the DEAE-cellulose chromatographic step of the purification protocol In fact, like for...
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Tài liệu Báo cáo Y học: Insights into the reaction mechanism of Escherichia coli agmatinase by site-directed mutagenesis and molecular modelling ppt
... one of the carboxylate oxygens of Asp153 and the carboxamide oxygen of Asn153 are positioned in such a way as to allow metal coordination interaction with one of the manganese ions This, together ... positions of other Fig Scheme of the binuclear manganese cluster and the localization of the side-chains of Asp153 and Asn153 in the modelled structures of wild-type and D153N mutant forms of E coli ... revealed by the conservation of the whole topology of the active site, would explain the essentially unaltered interaction of the D153N variant with the metallic cofactor In the modelled structure,...
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