use of site directed mutagenesis in the diagnosis prognosis and treatment of galactosemia
Allergic Diseases – Highlights in the Clinic, Mechanisms and Treatment Edited by Celso Pereira docx
... understanding of the interaction networks among humans, vaccines, drugs, and the environment and enable new insights of disease mechanisms and therapeutic responses The integration of all of the information ... work and interact with other systems The understanding of changes in molecular and cellular pathways and interactions can be useful for finding new drug, target and designing effective drugs These ... interest to further key points of clinical investigation Our understanding of the inflammatory process in allergic disease is clearly defined in many of these chapters, as is the application of science...
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ALLERGIC DISEASES – HIGHLIGHTS IN THE CLINIC, MECHANISMS AND TREATMENT potx
... understanding of the interaction networks among humans, vaccines, drugs, and the environment and enable new insights of disease mechanisms and therapeutic responses The integration of all of the information ... work and interact with other systems The understanding of changes in molecular and cellular pathways and interactions can be useful for finding new drug, target and designing effective drugs These ... interest to further key points of clinical investigation Our understanding of the inflammatory process in allergic disease is clearly defined in many of these chapters, as is the application of science...
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Báo cáo Y học: Role of tyrosine 238 in the active site of Rhodotorula gracilis D-amino acid oxidase A site-directed mutagenesis study docx
... in binding were observed for Y238 mutants with the ligands tested (Table 2) These results indicate that the mode of ligand binding is retained in the two mutants, and that the alteration of the ... exible and adapting their conformation depending on the size of the ligand side chain [27] Our results indicate that the role of Y223 and Y238 in the active site of RgDAAO is different from that of ... Y223 in binding and xation was suggested [4] This proposal was changed recently following the investigation of the effect of pH on benzoate binding in RgDAAO [12] and the resolution of the three-dimensional...
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Báo cáo khoa học: Probing the determinants of coenzyme specificity in Peptostreptococcus asaccharolyticus glutamate dehydrogenase by site-directed mutagenesis potx
... requires further structural insight is, on the one hand, the widespread use of Asp at the P7 position in many naturally occurring NADPH-dependent GDHs and, on the other hand, the failure of the E243D ... value in the absence of the other substrate In the presence of oxoglutarate, however, coenzyme binding was tighter and therefore measurable and, in particular, gave measurable Kd values for binding ... distinguish the cofactors in different ways [3] In the present study, we have investigated, by means of site- directed mutagenesis and steady-state kinetics, the individual contribution of the P7 glutamate...
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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
... located in the vicinity of the active site and could form a binding site for a glucosyl unit in subsite )1 (Fig 4B) Together with the observations from the site- directed mutagenesis, Trp402 of IPU ... for binding of Glc +2 of the a-(1fi4)-linked glucose units of pullulan (Fig 4A) IPU is an anomer-inverting enzyme Comparison of the active sites of the model of IPU and Dex49A The active site ... structure of the model of IPU N-terminal and b-helical domains are shown in light pink and grey, respectively The amino acid residues whose substitutions resulted in the reduction of the activity...
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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
... Subdomain I (N-terminal) contains the zinc-binding site, which faces into the deep cleft formed by the two subdomains connected at the base of the cleft The hinge-bending motion observed upon inhibitor ... binding to ACE2 (yellow) and tACE (white) (A) Binding site of CL1 in ACE2 and tACE; (B) binding site of CL2 in ACE2 and tACE Residue numbering for ACE2 is first The chloride ion is green and the ... conserved in ACE2 (His505) Indeed, the location (relative to the zinc-binding motif) of His505 in the ACE2 sequence is very similar to the location of the catalytic histidines in both ACE and thermolysin,...
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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
... Mn2+A in arginase I and Mn2+B in arginase II Because ligands to the metal ions are strictly conserved in these enzymes, the difference would reside in the length of the ligand– metal separations In ... agmatine hydrolysis, in studying the inhibitory effect of agmatine on arginine hydrolysis, reactions were followed by measuring the formation of ornithine, determined by the method of Chinard ... Differences in the 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...
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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
... which the enzyme was inactivated Binding of E1 to the peripheral subunit-binding domain of E2 The E1 component of the PDH complex of B stearothermophilus is bound to the E2 core mainly by interaction ... Temperature-dependence of the activity in the DCPIP assay of wild-type and mutant E1s E1b chains with the PSBD of the E2 chain To investigate whether this interaction was impaired by the mutations in the E1a ... for the cofactor ThDP to its binding site The lower thermal stability of the mutant E1s and the increased Vmax in the DCPIP assay, which could be due to easier access of DCPIP to the active site...
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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
... with the sequences of 1CEV and 1RLA and pasted into the structural alignment The alignment was then corrected by hand Since the sequences of the two arginases and agmatinase greately differ in the ... coordination interaction with one of the manganese ions This, together with the fact that the positions of other Fig Scheme of the binuclear manganese cluster and the localization of the side-chains ... amino acids at the beginning and the end, were mutated to glycines and, after remodelling back to their side chains in agmatinase, the whole hydrogen bond network was optimized The position of...
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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
... used to localize the active site (substrate-binding pocket) of AAO by molecular docking The enzyme consists of two domains, the FAD-binding domain (bottom part) and the substratebinding domain ... 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 ... group Other changes included introduction ⁄ removal of the phenolic hydroxyl in Tyr92 and Phe501, and substitution of His502 and His546 with leucine, serine and arginine residues Only three of the...
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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
... motifs in the binding site for HbpR and to examine the necessity of the sensing A-domain of HbpR for DNA binding The role of individual nucleotides and nucleotide motifs was characterized by site- directed ... important in forming the oligomeric structure and for DNA binding, by cloning an hbpR gene devoid of the A-domain, purifying this protein, and analyzing its DNA-binding characteristics and its ... of the hbpC promoter in Escherichia coli Results Mutagenesis of the HbpR-binding sites Sequence alignment of the UASs found in the HbpR operators in the hbpC and hbpD promoters defined a set of...
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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
... On the basis of the Ping Pong mechanism, the acetylation rate (k2) of NAT2 by the acetyl donor is independent of the transacetylation rate (k4) of the arylamine substrate Because the values of ... Tyr190, the side chain of Asn72 and the backbone of Gly124 and Ala123 participate in a network of interactions with Asp122 Moreover, because the centroid of the Tyr190 phenyl ring is $ 3.5 A from the ... stabilizes the protonated His, increasing its pKa value and, therefore, increasing the nucleophilicity of active site Cys A similar interaction was found between the indole ring of an active site Trp177...
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Báo cáo khoa học: Residues affecting the chloride regulation and substrate selectivity of the angiotensin-converting enzymes (ACE and ACE2) identified by site-directed mutagenesis pot
... tACE and ACE2 The residues surrounding the chloride ion at the CL1 and CL2 sites of tACE and ACE2 are shown in Fig In tACE, the residues that coordinate the chloride ion at the CL1 site are Arg186, ... stabilizing complex formation for the residues in subdomain II involved directly in substrate binding [20] The CL2 site of ACE has previously been suggested to contain the mechanistically binding ... R514 that has the greatest in uence on the chloride regulation of activity, and that the CL2 site of ACE does indeed contain the mechanistically binding chloride ion The ACE2 CL2 site residue...
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Báo cáo khoa học: Investigation of the substrate specificity of a b-glycosidase from Spodoptera frugiperda using site-directed mutagenesis and bioenergetics analysis pdf
... Bioscience) The nonretained proteins were washed out in the dialysis buffer, while the retained proteins were eluted from the column using an NaCl gradient prepared in the same buffer The presence of the ... independent and also compares the energy of ESà complexes containing different sets of noncovalent interactions between E and Sà The steps used to calculate the energy of individual noncovalent interactions ... (dotted lines) involving the residue 39 and E451 were represented 4-OH is represented in the equatorial (4e; pointing towards left) and axial (4a; pointing upwards) positions 3-OH is always in the...
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Báo cáo khoa học: Probing the unfolding region of ribonuclease A by site-directed mutagenesis potx
... acceleration of the unfolding reaction, i.e kinetic destabilization, for the variants and the similarity of DDG[D]50% and DDG#U indicates that the decrease in the thermodynamic stability is mainly caused ... an increase in kU Furthermore, our results suggest that Leu35, the side chain of which is buried in the interior of the molecule like that of Phe46 (Table 2), is involved in the formation of ... introduction of the proline residues, i.e the decrease in loop exibility, did not increase the global stability of the RNase A molecule By introducing a cleavage site for thermolysin (S21LRNase A), the...
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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
... when the crystal structure of the spinach enzyme was determined, first in the presence of the inhibitor IpOHA [8] and later with product bound in the active site [9] There are several interesting ... from that of the spinach enzyme with the active site constructed from two monomers of a dodecamer In contrast, the active site of spinach KARI is wholly contained within each monomer of a tetramer ... participates in the isomerase reaction while E319 is involved in the reductase However, it is also possible Table Corresponding active site residues of KARI of spinach, P aeruginosa and E coli In the...
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Báo cáo khoa học: Mapping of the active site of glutamate carboxypeptidase II by site-directed mutagenesis potx
... distortion of the coordination sphere of the active -site zincs Our results indicate that the binding of the glutarate part of the inhibitor in the S1¢ pocket contributes to the inhibition effect of the ... mutations of amino acids delineating the substrate binding cavity of GCPII were designed and introduced into the GCPII ectodomain (rhGCPII; amino acids 44–750) using site- directed mutagenesis Individual ... productive architecture of the S1¢ site of GCPII, including the positioning of the Tyr552 side chain Despite similarities between lysine and arginine residues, the lysine side chain could not fully...
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Báo cáo khoa học: Site-directed mutagenesis and footprinting analysis of the interaction of the sunflower KNOX protein HAKN1 with DNA ppt
... right of each panel indicate the DNA sequence (5¢ end in the upper part) of the corresponding strand in this region Below the footprints, the sequence of the corresponding binding site is shown and ... both binding sites in the complexes formed by KNOX and Bell proteins Sequences outside the homeodomain may also in uence the binding properties of the protein Indeed, a stretch of 16 amino acids ... Analysis of DNA binding by hydroxyl radical footprinting and interference assays A more detailed picture of the binding of HAKN1 to its target site was obtained by the analysis of footprinting patterns...
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Báo cáo khoa học: Defining the QP-site of Escherichia coli fumarate reductase by site-directed mutagenesis, fluorescence quench titrations and EPR spectroscopy doc
... between the heme and the [3Fe-4S] cluster of SdhB [1,6] In the case of FrdABCD, the membrane-intrinsic domain does not contain heme, but instead contains two menaquinones at discreet sites in the ... would therefore be of interest to examine the effects of a range of sitedirected mutants on the HOQNO binding properties and enzymology of the enzyme In this paper, we evaluate the effects of mutation ... QP site (the proximal Q -site) , is located in the interface region between the FrdCD subunits and the [3Fe-4S] cluster coordinating region of FrdB on the cytoplasmic side of the membrane The other...
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Báo cáo Y học: Potential active-site residues in polyneuridine aldehyde esterase, a central enzyme of indole alkaloid biosynthesis, by modelling and site-directed mutagenesis ppt
... Nevertheless, the specific activities of each of the muteins decreased With the exchange of Cys132, there was a dramatic decrease (fourfold) in the specific activity, and in the case of Cys170, the ... cysteine and serine protease inhibitors The next clear information obtained was the necessity of histidine groups which proved the active role of at least one histidine in substrate binding The following ... excess of this mutein gave no Fig Stereo pictures of the modelled structure of (His)6PNAE Only the backbone and the side-chains of the mutated amino acids are shown The amino acids of the catalytic...
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