biological activity of insecticidal toxins structural basis site directed mutagenesis and pers
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
... 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 structures of the model of IPU (Fig 4A) and Dex49A ... catalytic residues of IPU Also, Trp425 of Dex49A, which is the corresponding residue of Trp402 of IPU, is reportedly located in the vicinity of the active site and could form a binding site for a glucosyl ... potential subsites )1 and )2 are reportedly more conserved in GH family 49 than the residues in subsites +1 and +2 [17] Comparison of the active sites of the model of IPU and Dex49A clearly shows...
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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
... ligand-manganese distances RESULTS AND DISCUSSION By site- directed mutagenesis, a D153N mutant form of E coli agmatinase was obtained The mutant enzyme retained about 5% of wild-type activity and ... 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 ... modeling of the active site The modelled structure was very similar to the template, with respect to the number and arrangement of structural elements (a-helix and b-sheets), and one of the major...
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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
... study, the interaction of human arginase II with substrate and metal ions was examined by sitedirected mutagenesis and kinetic studies Selection of target residues (Asp149, His120 and His145) was based ... Interaction of arginase II with substrate and manganese ions tional evidence for the differences between the active sites of this enzyme and arginase I In spite of the relatively low agmatinase activity ... corresponding His101 and His126 in arginase I, were described among the ligands for coordination of Mn2+A and Mn2+B, respectively [9,24] However, the volume of the active site cleft was found...
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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
... the specificity of new Sfbgly50 mutants Materials and methods All reagents, unless otherwise specified, were purchased from Sigma or Merck Site- directed mutagenesis Site- directed mutagenesis was ... confirmed by SDS/PAGE This procedure yielded % 0.5 mg of mutants E451S and E451Q, 0.3 mg of mutant Q39E and 0.15 mg of mutants Q39N and E451D, from 0.5 L of bacterial culture Steady-state kinetic parameters ... corresponding to energy of the hydrogen bond between residue 39 3-OH (/3), which was added to the energy of /4e /4a interactions was the and and Results and Discussion The induction of recombinant enzymes...
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Báo cáo khoa học: Mapping of the active site of glutamate carboxypeptidase II by site-directed mutagenesis potx
... Sweeden) and 3H-NAAG substrate was obtained from Perkin-Elmer (Boston, MA, USA) Site- directed mutagenesis Site- directed mutagenesis was carried out using the QuikChange Site- Directed Mutagenesis ... decrease of both the Km and kcat values The mutation of Gly518 to Pro led to a slight increase of the Km value and one order of magnitude decrease of the kcat value (Table 2) Analysis of inhibitor ... pocket of human GCPII using molecular modeling and sitedirected mutagenesis analysis Guided by the previously determined crystal structure of GCPII, we set out to complement the available structural...
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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
... expression and purification of recombinant proteins HAKN1 homeodomain and C-terminal sequences were amplified and cloned in-frame into the EcoRI and PstI sites of the expression vector pMAL-c2 (New England ... panels in (A) and (B) represent the top and bottom strands of the binding site, respectively A portion of the same fragment digested with defined restriction enzymes was used as a standard to calculate ... incorporation of Ile50 and Arg55 must have been particularly important Other homeodomains that contain Ile50 and Arg55 are those of the TGIF, Meis and Bell families [22] and yeast MATa1 [36] TGIF and Meis...
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Báo cáo khoa học: The phosphate site of trehalose phosphorylase from Schizophyllum communeprobed by site-directed mutagenesis and chemical rescue studies pptx
... C Goedl and B Nidetzky The phosphate site of Schizophyllum commune trehalose phosphorylase scanning mutagenesis of the active site of trehalose phosphorylase [16] and report on results of steadystate ... mm) and K512A by guanidine (10200 mm) was observed Addition of 200 mm of NaCl did not alter the activity of either one of the site- directed mutants These results Fig Functional complementation of ... of the pKa of the amine and guanidine derivatives on specic restoration of activity in K512A and R507A, respectively, was very small and probably not signicant Analysis of the pH dependence of...
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Tài liệu Báo cáo khoa học: Design, structure and biological activity of b-turn peptides of CD2 protein for inhibition of T-cell adhesion ppt
... bonds between NH of Thr5 and CO of Arg2, as well as NH of Arg2 and CO of Thr5 Structure of cyclic hexapeptides – cYT peptide The NMR data of the cYT peptide were indicative of its flexible nature ... biological activity and to study the effect of reducing the chain length of peptides on Ó FEBS 2004 Design of peptides for T-cell adhesion inhibition (Eur J Biochem 271) 2879 biological activity, ... intensity of ROE cross-peaks, and were assigned as strong, medium and weak ˚ represent the family Based on ROE violation > 0.2 A and allowed values of /, w in the Ramachandran map, only one family of...
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Tài liệu Báo cáo khoa học: Physicochemical characterization and biological activity of a glycoglycerolipid from Mycoplasma fermentans ppt
... presence of minute amounts of endotoxins The comparison of LPS and MfGl-II in the LAL assay shows, as expected, activity of LPS in the range of down to 10 pgÆmL)1, whereas MfGl-II has only activity ... Schromm, A.B., Koch, M.H.J & Seydel, U (1995) Conformation and fluidity of endotoxins as determinants of biological activity In Bacterial Endotoxins: Lipopolysaccharides from Genes to Therapy (Levin, ... water-binding capacity of lecithin headgroups [30] Finally, MfGl-II exhibits a main band at 1245 cm)1 and further weak bands at 1220 and 1260 cm)1 Thus, the phosphate groups of this glycolipid are...
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Tài liệu Báo cáo khoa học: Moult cycle-related changes in biological activity of moult-inhibiting hormone (MIH) and crustacean hyperglycaemic hormone (CHH) in the crab, Carcinus maenas From target to transcript ppt
... (or both) of the above-mentioned processes may be relevant to moult control, we first investigated the biological activity of MIH and CHH during precisely timed stages of the moult cycle of Carcinus ... MIH content of the SG, relative to CHH, during late premoult Expression patterns of MIH and CHH in the X-organ during the moult cycle Expression patterns of mRNA levels of both MIH and CHH were ... investigated levels of MIH and CHH peptides in the sinus gland and levels of transcripts in XO neurones during the moult cycle, to see whether significant events, such as up- or down-regulation of transcription...
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Báo cáo khoa học: Construction and biological activity of a full-length molecular clone of human Torque teno virus (TTV) genotype 6 pptx
... [19], bone marrow [10] and several other organs [20] However, the main host cells and the target organs of this virus are still undefined The study of the biological functions of TTV is particularly ... interactions, and to find permissive cell lines for further molecular and cell biological studies of this peculiar human virus For this purpose, we have cloned and sequenced the full-length genome of TTV ... SalI double-digestions of the input linear construct yielded three restriction fragments of approximately 2100, 740 and 890 bp with the first enzyme pair, and of 2200, 630 and 890 bp with the latter...
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Báo cáo khoa học: Natural and amyloid self-assembly of S100 proteins: structural basis of functional diversity doc
... response and a factor in host ⁄ parasite defences, which binds Cu2+ and Zn2+ at the same site and corresponds to the Zn2+-binding site in S100A7, evoking a possibly similar structural and functional ... higher level of understanding the biology of tumour and degeneration and enable to use our acquired knowledge of S100 structure and functions in developing strategies to modulate their activity for ... the EF-hands in S100 proteins Structure of the N-terminal, S100-specific EF-hand (A) and the C-terminal, canonical EF-hand (B) in the metal-free (lighter) and Ca2+-bound (darker) form of S100A6...
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Báo cáo khoa học: Enhancement of intracellular concentration and biological activity of PNA after conjugation with a cell-penetrating synthetic model peptide docx
... significantly augments the biological activity of a 12-mer antisense PNA directed against the nociceptin/orphanin FQ receptor Pretreatment of CM with 0.2 lM of the naked PNA (I; Table 1) and its MAP-conjugate ... influence of nociception on the beating rate of CM measured on day of culture without and with pretreatment with either 0.2 lM of I or II on day and after seeding The basal beating rate of the CM ... down-regulation of the nociceptin/orphanin FQ receptor to be the mechanism of biological activity of compounds I and II The antisense pretreatment remained without any influence on the basal beating rate of...
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Báo cáo Y học: Synthesis, conformational analysis and biological activity of cyclic analogs of the octadecaneuropeptide ODN Design of a potent endozepine antagonist pot
... analysis of the calculated structures revealed a short distances between NH of Leu7 and the side chain carboxyl group of the Asp6 residue (< 0.28 nm), and NH of Leu7 residue and C¼O of Leu5 (< ... of a potent endozepine antagonist (Eur J Biochem 268) 6049 Fig RP-HPLC monitoring of N- and C-terminal deprotections and on-resin cyclization of the linear precursor of cyclo128 OP Aliquots of ... the receptor affinity and selectivity of peptide ligands [13,14,27] The Ala-scan of OP has revealed that the side chain of each residue is required for the full activity of the peptide [12] These...
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báo cáo khoa học: " Biological activity of a genetically modified BMP-2 variant with inhibitory activity" potx
... in vitro evaluation of their biological activity, using ALP activity as a marker, revealed alterated effects for mutants of epitope and epitope as well But only alterations of epitope lead to ... opposite limb Three groups with individuals each were established, using doses of 10, 40 and 160 μg Thus the number of animals was n = 18 After a period of 28 days the animals were sacrificed and ... heterotopic implantation site (skeletal muscle) and BMP-2 wild type in a well known dose as an agonistic stimulus was chosen Methods Origin of the proteins The developement and expression of the utilized...
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Tài liệu Báo cáo khoa học: The ubiquitin ligase Itch mediates the antiapoptotic activity of epidermal growth factor by promoting the ubiquitylation and degradation of the truncated C-terminal portion of Bid ppt
... between EGF-induced reduction in Bid and tBid levels and the ubiquitin ligase activity of Itch In this study, we first examined the ability of Itch to interact with Bid and tBid We found that Itch specifically ... inset) n = conditions [29] Treatment of HEK-293T cells with TRAIL is known to induce caspase activity and cleavage of Bid in tBid [30] In our hands, treatment of HEK-293T cells with 200 ngÆmL)1 ... ± 0,02% of control; P = 0.001 for cell survival and 0.94 ± 0.03 of control for caspase activity) , and treatment with EGF slightly increased this effect on cell survival (88.4 ± 2.0% of control;...
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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
... 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 ... activity was investigated (Fig 4) The hydrolysis of Mca- Fig Role of His505 and His345 in catalysis Schematic of the proposed reaction intermediate of ACE2, showing the importance of His345 and ... ion-binding sites of each enzyme The designated CL2 site in tACE is absent from ACE2 [20], and this is due to the substitution of Pro407 and Pro519 in ACE for Glu398 and Ser511 in ACE2 The side chains of...
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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
... motif at the active site is underlined P1 and P2 mark phosphorylation sites and (serine residues) in the E1a chains of eukaryotic PDH and BCDH complexes ODPA and ODPT, E1a chain of PDH complexes ... PLOT software 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 ... two active sites of pigeon breast muscle E1p work in an Ôalternating site mechanismÕ and that intersubunit interactions may play an essential part in the catalysis by, and regulation of, the enzyme...
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Tài liệu Báo cáo Y học: Study of substrate specificity of human aromatase by site directed mutagenesis pdf
... (c) the steric constraints of the K130-F134 complex to the C(19) methyl of testosterone; and (d) the role of D309 and of E302 in aromatization of androgens and nor-androgens, respectively Thus, ... on the basis of the atomic partial lipophilicity values [44] and a distance-dependent function [45] Site- directed mutagenesis This step was performed with the QuickChangeTM SiteDirected Mutagenesis ... substrate and inhibitors Docking of the androstenedione into the active site was carried out considering the orientation of C(1), C(2) and C(19) above the heme [41] and the position of the ligand towards...
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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
... above A Site- directed mutagenesis of aryl-alcohol oxidase model, site- directed mutagenesis and kinetic studies were used to identify the enzyme active site and evaluate the role of some selected residues ... et al Site- directed mutagenesis of aryl-alcohol oxidase H546 A B H502 N732 C Fig Conserved residues at the active site of four GMC oxidoreductases The positions of conserved histidine and histidine ... side chains of Tyr78, Tyr92, Leu315 and Phe501 to a methyl group Other changes included introduction ⁄ removal of the phenolic hydroxyl in Tyr92 and Phe501, and substitution of His502 and His546...
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