directed mutagenesis in structure activity studies of neurotransmitter transporters
Báo cáo khoa học: Site-directed mutagenesis, kinetic and inhibition studies of aspartate ammonia lyase from Bacillus sp. YM55-1 pptx
... effects, including the suboptimal positioning of the amino group and the slower release of product In the substrate binding model of AspB, two other residues (Thr101 and Asn142) are implicated in binding ... role for the lysine residue in binding one of the two carboxylate groups of the substrate According to the substrate-binding model of AspB, Lys324 binds the C1 carboxylate group of [23] The strongest ... upon substrate binding, positioning Ser318 in the vicinity of C3 of the substrate, we have initiated studies that aim to solve the X-ray structure of the enzymesubstrate complex.) In the next step,...
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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
... phosphorylases A plot of concentration of G1P released against the incubation time was linear in all cases, indicating that enzyme inactivation did not interfere with determination of the initial rate ... phosphorylase activity in R507A and K512A Inclusion of 200 mm of guanidine into the assay for phosphorolysis of a,a-trehalose at pH 6.6 caused 45-fold enhancement of the activity of R507A seen in the ... experiments, in which a small molecule compensates for the missing side chain of a relevant site -directed mutant, often provide valuable insights into the role of active-site arginine [3236] and lysine...
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Báo cáo khoa học: Structure–activity relationships of fowlicidin-1, a cathelicidin antimicrobial peptide in chicken docx
... Results Solution structure of fowlicidin-1 To determine the secondary structure of fowlicidin-1, CD spectroscopy was performed in increasing concentrations of the structure- promoting agents trifluoroethanol ... FEBS Y Xiao et al Structure activity relationships of fowlicidin-1 LPS-binding sites [16] To map the regions involved in the binding of fowlicidin-1 to LPS, the N- and C-terminal deletion mutants ... C-terminal end [25] To determine whether the two LPS-binding sites act in a synergistic manner, an equimolar mixture of fowlicidin-1(1–15) and fowlicidin-1(8–26), each containing one LPS-binding...
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Tài liệu Báo cáo khoa học: Structure-activity relationships of a-conotoxins targeting neuronal nicotinic acetylcholine receptors ppt
... of the structure also contain hydrogen bonds Several structures have been determined independently by different groups and the structures appear to be in good agreement The structure of ImI in ... capping (or initiating) the helical element these residues may play a vital role in correctly orienting key binding residues Arg7 was suggested to have a functional role in binding as the structure ... three-dimensional structures of the mutants were determined [21,46] It was noted that very small conformational changes in ImI, especially for the side chains involved in binding, are associated with a loss of...
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Tài liệu Báo cáo khoa học: Second messenger function and the structure–activity relationship of cyclic adenosine diphosphoribose (cADPR) doc
... N1 of inosine (Fig 4), while N7-cIDPR showed no Ca2+ release activity in sea urchin egg homogenates [59] Indeed, Ca2+ release activity of inosine derivatives was first described for a series of ... cyclized between N1 of inosine and C2 of the northern ribose [60] In addition, 8-Br-N1-cIDPR induced Ca2+ signalling in 4594 intact T-cells [61,62] This finding was surprising since 8-Br-N1-cADPR ... direct binding site of cADPR at the RyR has not been described so far In contrast, in a limited number of cell systems FK506 binding protein 12.6 (FKBP 12.6, calstabin2) was found to bind cADPR...
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Báo cáo khoa học: Structural diversity of angiotensin-converting enzyme Insights from structure–activity comparisons of two Drosophila enzymes docx
... kcat, and replacing the Phe8 of BK with tyrosine resulted in a modest increase in both the Km and kcat In contrast, replacing Ser6 of BK with threonine resulted )1) in greatly increased affinity ... FEBS R J Bingham et al inal domain of human angiotensin I-converting enzyme: Insights into the origins of its exopeptidase activity J Biol Chem 280, 6669–6675 34 Riviere G, Michaud A, Deloffre L, ... R J Bingham et al Structure- activity of Drosophila ACEs Table Effect of NaCl on the kinetic constants for the conversion of AI into AII by ANCE Kinetic constants for the conversion of AI into...
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Model choice in time series studies of air pollution and mortality pdf
... decrease in the bias of β with increasing df is explained in Dominici et al (2004) for the natural spline case and explored by Rice (1986) and Speckman (1988) in the nonparametric setting Dominici ... sensitivity of β by varying the df that is used in the model(s) and examining the associated ˆ changes in β In summary, we explore the following strategies for deciding on an appropriate number of degrees ... notion of ‘degrees of freedom’ ∼ N 0, σ / σ > 2.1 Representing f Common choices for representing the smooth function f in model (1) include natural splines, penalized splines and smoothing splines...
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Báo cáo sinh học: "Paracentric inversion involving NOR of chromosome 8 in a boar : studies of synaptonemal complexes under a light microscope" pdf
... been no report concerning any case of inversion in this species The existence of an inversion in a karyotype can be a cause of pairing disturbances at the pachytene substage of meiotic prophase ... to crossing over within the loop and, in consequence, to the production of unbalanced gametes On the other hand, lack of loop formation results in supression of crossing over within the inverted ... by application of conventional and banding stainings showed in chromosome No an additional chromatin fragment within the nucleolar organizer region (NOR) Silver staining revealed in this chromosome...
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Structure function studies of vesicle associated membrane protein associated protein b (VAPB) associated with amyotrophic lateral sclerosis (ALS
... properties of the wt-VAPB MSP domain 39 3.1.2 Stability of the wt-VAPB MSP domain 41 3.1.3 Binding activity of the wt-VAPB MSP domain 41 3.1.4 Crystal structure of the wt-VAPB MSP domain 42 Studies ... Interactions with FFAT-motif containing proteins The hVAP proteins have been shown to interact with proteins carrying a short motif consisting of two phenylalanines in an acidic tract (Wyles et al., ... presence of hVAP MSP domain in the blood serum using mass spectrometry Tsuda et al., (2008) also showed that the hVAP MSP domain is able to bind the extracellular ligand binding domain of EphA4 in...
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Prevalence and trends in overweight and obesity in threecross sectional studies of British children, 1974-94
... Rising trends in children will almost certainly be represented in later trends in adult overweight and obesity and probably in an increase in associated adult morbidity We thank our colleagues in ... Body mass index in children worldwide: cut-off points for overweight and obesity BMJ 2000;320:1240-3 Chinn S, Hughes JM, Rona RJ Trends in growth and obesity in ethnic groups in Britain Arch Dis ... particularly noticeable in Scottish children.8 Although debate over the cut-off points will continue, this should not detract from the urgency of tackling the problem of obesity Our data indicate that...
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ANALYSIS OF MICROBIAL COMMUNITY STRUCTURE AND IN SITU ACTIVITY OF NITRIFYING BIOFILMS
... distributions of in situ nitrifying activities were determined The relationship between the spatial organization of nitrifying bacterial populations and the in situ activity of these populations within ... microprofiles of NH4+, NO2-, and NO3- in an autotrophic nitrifying biofilm that was initially fed with enrichment culture of nitrifying bacteria (bioaugmented biofilm) This bioaugmented biofilm ... were performed In situ hybridization of vertical biofilm thin sections clearly indicated that the numbers of probe NEU-stained NH4+-oxidizing bacteria (i.e., Nitrosomonas marina-lineage, Nitrosomonas...
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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
... zinc protease domain is divided into two subdomains, which are defined by the movement of the subdomains relative to each other upon inhibitor binding Subdomain I (N-terminal) contains the zinc-binding ... subdomains [20] Subdomain I contains the zinc ion and the N-terminus The C-terminus is found in subdomain II Enzyme activity, % of activity at 0.5 M NaCl 100 80 tACE 60 Subdomain I 40 Subdomain II ... C-terminus of the ACE2 inhibitor, MLN-4760 [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...
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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
... between arginine and agmatine Agmatine, which results from decarboxylation of arginine by arginine decarboxylase, is a metabolic intermediate in the biosynthesis of putrescine and higher polyamines ... 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 ... by agmatine in E coli agmatinase and l-arginine in B caldovelox arginase [37], indicating that the substrate specificity of these enzymes rely mainly in substituents at C-a This has been, in fact,...
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Tài liệu Báo cáo Y học: A Raman optical activity study of rheomorphism in caseins, synucleins and tau New insight into the structure and behaviour of natively unfolded proteins pot
... accumulating that this originates in a more hydrated form of b strand [18] The positive bands at » 1675 cm)1 in the amide I region of the ROA spectra of b- and j-casein, which originate mainly in the ... observed in the caseins [3], synucleins [10], tau [14], prothymosin a [7] and the ®bronectin-binding protein [8], and also in typical chemically denatured proteins [56±58] Over extensive lengths of ... above In view of the close similarity of the ROA spectra of the casein, synuclein and tau proteins shown in this paper, we have reproduced in Fig from an earlier study [22], the ROA spectrum of...
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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
... was changed recently following the investigation of the effect of pH on benzoate binding in RgDAAO [12] and the resolution of the three-dimensional structure of RgDAAO in complex with anthranilate ... (retaining their FAD prosthetic group) The mutants, in their oxidized state, show the typical spectrum of the FADcontaining avoproteins (line in Fig 2), an extinction coefcient at 455 nm of % ... band at % 600 nm following the binding of anthranilate (Fig 3, inset) and the shoulder at % 500 nm following the binding of benzoate (De497nm of 7500ặM)1ặcm)1 vs a gure of 2000 4000ặM)1ặcm)1 observed...
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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
... hydrolytic activity of PNAE In order to gain evidence for these suggestions, inhibitor studies, site -directed mutagenesis experiments and modelling of PNAE were performed Inhibitor studies Several ... regarding studies with 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 ... quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal Biochem 72, 248–254 15 Sali, A & Blundell, T (1993) Comparative protein modelling by satisfaction of...
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Báo cáo khoa học: Probing the determinants of coenzyme specificity in Peptostreptococcus asaccharolyticus glutamate dehydrogenase by site-directed mutagenesis potx
... evolution of nucleotide-binding protein Nature 250, 194–199 Wierenga RK, Terpstra P & Hol WG (1986) Prediction of the occurrence of the ADP-binding bab-fold in proteins, using an amino acid sequence ... However, in this latter case, the interaction of enzyme and coenzyme caused a very small change in fluorescence The measurements with NADH in the absence of oxoglutarate show a weakening of 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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Báo cáo khoa học: Tracking interactions that stabilize the dimer structure of starch phosphorylase from Corynebacterium callunae Roles of Arg234 and Arg242 revealed by sequence analysis and site-directed mutagenesis doc
... destabilizing in unligated StP, site -directed mutagenesis of the side chain of Arg234 into the methyl side chain of alanine did not diminish the stabilizing effect of sulfate binding in comparison ... replacement of the guanidinium side chain of arginine by a methyl side chain of alanine in the R234A and R242A mutants caused only a small effect on the binding of sulfate An approximately 2.5-fold increase ... measured kinetic stabilities Arginine residues are known for their prevalence in both intra- and inter-chain interfaces [22,23] where the charged guanidinium group is often involved in formation of...
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Aerobic Uptake of Cholesterol by Ergosterol Auxotrophic Strains in Candidaglabrata & Random and Site-Directed Mutagenesis of ERG25 in Saccharomyces cerevisiae
... starting temperature of 195˚C for one minute increasing to 240˚C in 20˚C/min increments and then 2˚C/min increments to a final temperature of 280˚C where the temperature held for five minutes ... 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 Candida ... was used with a starting temperature of 100˚C for one minute increasing to 300˚C in 7˚C/min increments and then held for 15 minutes at the final temperature of 300˚C The injection volume per...
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Mechanistic studies of anti malarial spiroindolones and synthesis and structure activity relationship studies of an inhibitor of dengue proliferation
... the role of the imine configuration in governing the final stereochemistry of the indoline-spirotetrahydro-β-carboline structure shown in Figure 10 remained uncertain In order to obtain a better ... combination of sulfadoxine and pyrimethamine is also commonly employed in the treatment of malaria in pregnant women And in the event of severe multi-drug resistant malaria, quinine taken in combination ... THE DEGREE OF MASTER OF INFECTIOUS DISEASES, VACCINOLOGY AND DRUG DISCOVERY YONG LOO LIN SCHOOL OF MEDICINE NATIONAL UNIVERISTY OF SINGAPORE AND SWISS TROPICAL INSTITUTE UNIVERSITY OF BASEL Acknowledgements...
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