metalloproteins and metal binding sites
Báo cáo khóa học: Mutational and structural analysis of cobalt-containing nitrile hydratase on substrate and metal binding pdf
... DNA polymerase (Toyobo, Japan), using the bF and 68r primers, bR and 68f primers, aF and 109r primers, aR and 109f primers, aF and 114r primers, or aR and 114f primers Following an initial denaturation ... 55 °C, and a incubation at 68 °C The second PCR was also performed with KOD-plus DNA polymerase, using the two megaprimers and the bF and bR primers, or the two megaprimers and the aF and aR ... reaction, was used The 0.72 kb DNA fragment and pUC18-NHase were digested with EcoRI and NotI, and the 1.08 kb DNA fragment and pUC18-NHase were digested with NotI and HindIII The PCR product was ligated...
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Báo cáo khoa học: Co-operation of domain-binding and calcium-binding sites in the activation of gelsolin pptx
... 1.5 lM, and another around 0.1 lM These transitions reflecting conformational changes correlate well with the two binding sites (IIG5 and IIG6, Kd ¼ and 0.2 lM, respectively) detected and measured ... hydrophobic and salt bridges between G2 helix a1 and the C-terminal helix of G6 It is proposed that binding of IIG6 disrupts the salt bridges between D669 and R168, and between D670 and R169 Cooperativity ... dissociation constant Kd and the maximal binding Amax) were determined by nonlinear fitting A ¼ Amax · [L]/(Kd + [L]) (relation 1) where A is the absorbance at 405 nm and [L] the ligand concentration,...
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BIOCHEMICAL APPLICATIONS OF DSRED-MONOMER UTLIZING FLUORESCENCE AND METAL-BINDING AFFINITY
... metal- binding sites within the DsRed-Monomer Sequence identity comparison with known metal- binding proteins is a common method of locating metal- binding sites within a protein or enzyme MetalMine ... identity and position of possible metal binding motifs within the structure and sequence of DsRed-Monomer, two computational programs were utilized MetalMine and Metsite were used to help locate metal- binding ... MetalMine compares the target protein sequence with a compiled list of metal binding proteins and enzymes Metsite looks for metal binding sites within the sequence computationally MetSite uses a set of...
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Identification of the Pba1 and Pba2 Binding Sites on 20S Core Particle Intermediates
... proteasomal binding sites, having been successfully used to identify binding sites for other HbYX-‐motif-‐containing proteins; further assay optimization should yield Pba1p and ... University, August 2012 Identification of the Pba1 and Pba2 Binding Sites on 20S Core Particle Intermediates Major Professor: Andrew Kusmierczyk The proteasome is responsible ... UNIVERSITY GRADUATE SCHOOL Research Integrity and Copyright Disclaimer Title of Thesis/Dissertation: Identification of the Pba1 and Pba2 Binding Sites on 20S Core Particle Intermediates For the...
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Tài liệu Báo cáo Y học: Interaction of decorin with CNBr peptides from collagens I and II Evidence for multiple binding sites and essential lysyl residues in collagen pot
... conclude the precise location and the relative orientation of the binding sites in decorin and collagen are not yet known Our findings on multiple binding sites in collagen and on the relevance of Lys/Hyl ... decorins and decorin cores, respectively Left lane: standard protein markers and their molecular masses (in kDa) The core protein is present as two bands with apparent molecular masses of 47 and 42 ... interactions between type I and type II collagens, their peptides and decorin reveal the following (a) Type I and probably also type II collagen appear to have multiple binding sites for decorin, because...
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Tài liệu Báo cáo Y học: Short peptides are not reliable models of thermodynamic and kinetic properties of the N-terminal metal binding site in serum albumin doc
... of Cu(II) and Ni(II) exchange by albumin, we characterized the binding parameters and performed parallel kinetic studies using HSA and BSA and three simple analogues of the N-terminal binding site ... varied metal ion amounts The albumin samples for titrations and metal exchange kinetics measurements were kept at pH 7.4 (100 mM sodium phosphate buffer) The kinetics of metal binding to peptides and ... considered Comparison of Cu(II) and Ni(II) binding between model peptides and albumins Afnity for Ni(II) at site I can be compared between albumins on one hand and DAHam and DAHKam on the other Much...
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Báo cáo khoa học: Ion-binding properties of Calnuc, Ca2+ versus Mg2+ – Calnuc adopts additional and unusual Ca2+-binding sites upon interaction with G-protein pdf
... Ca2+ -binding sites is known to generate the J-band (610–650 nm) and ⁄ or the c-band (480–510 nm) Upon binding to Calnuc, the dye displayed prominent J-band and c-band in both absorption and CD ... the J-band and the c-band regions (Fig 6B) on binding to the protein CD results confirmed the absorbance data: Ca2+ binding is able to attenuate both the J-band and the c-band, whereas Mg2+ binding ... b-crystallin and d-crystallin behave similarly (show only one band), whereas calmodulin and troponin behave like each other On the other hand, Calnuc and parvalbumin generate both the J-band and the c-band,...
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Báo cáo khoa học: Caenorhabditis elegans metallothionein isoform specificity – metal binding abilities and the role of histidine in CeMT1 and CeMT2 potx
... and CeMT2 metallothioneins Fig Comparison between the CD spectra of recombinant CeMT1 and CeMT2 (black), NtCeMT1 and NtCeMT2 (red) and CtCeMT1 and CtCeMT2 (green) synthesized in Zn- and Cd-supplemented ... number of CeMT1 and CeMT2 histidines contributing to divalent metal ion coordination, the Zn and Cd preparations of both C elegans CeMT1 and CeMT2, and the Cd complexes of CtCeMT1 and CtCeMT2, were ... Call2 and CalliCu, Callinectes sapidus, 1, and Cu isoforms (Uniprot accession numbers Q548Y3, Q548Y2 and Q9U620, respectively); Scy1 and Scy2, Scylla serrata isoforms and (Uniprot P02805 and P02806);...
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Báo cáo khoa học: Expression of the Drosophila melanogaster ATP synthase a subunit gene is regulated by a transcriptional element containing GAF and Adf-1 binding sites pptx
... was considered as 100%) was mutated and the activity of the mutant constructs was assessed Wild type and mutated GAGA and Adf-1 binding sites are shown by open and filled symbols, respectively (C) ... region of the a-F1-ATPase gene and found that the combination of GAF and Adf-1 binding sites is present both in other Drosophila species (D yakuba and D pseudoobscura) and in Anopheles gambiae (data ... singlestranded binding protein (mtSSB), and the catalytic (a) and accessory (b) subunits of the DNA polymerase c (Pol c) [24,25] Interestingly, the expression of the genes encoding mtSSB and Pol...
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Báo cáo khoa học: Carbohydrate binding sites in Candida albicans exo-b-1,3-glucanase and the role of the Phe-Phe ‘clamp’ at the active site entrance ppt
... from sugar binding at the +2 and +3 subsites FEBS Journal 277 (2010) 4549–4561 ª 2010 The Authors Journal compilation ª 2010 FEBS W M Patrick et al Carbohydrate binding sites in Candida exoglucanase ... of L3 binding in the active site pocket showing the Phe-Phe clamp and the F229A mutation Carbohydrate binding subsites are labelled )1, +1 and +2 Glycosidic bond cleavage occurs between )1 and ... Exg and a carbohydrate binding module CBM 4-2 from T maritima (PDB: 1gui) is shown with F144, F258 and F229 from Exg (cyan), and W51, W102 and W27 from CBM 4-2 (magenta) from the associated binding...
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Báo cáo Y học: SMAP-29 has two LPS-binding sites and a central hinge docx
... contained multiple binding sites and its LPS binding manifested positive cooperativity Should a Ôtake-home messageÕ be needed from the present study, one might remember Gulliver and the Lilliputians ... to SMAP-29 [9–29] suggests that the Nterminal binding site has higher affinity for LPS than does the C-terminal one The presence of two binding sites and the sevenfold to 21-fold greater affinity ... therefore infer that cooperativity in LPS -binding is primarily intramolecular, whereas binding of one LPS -binding site in a SMAP-29 molecule facilitates binding by the other site The Hill coefficient...
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Báo cáo khoa học: Polysaccharide binding sites in hyaluronate lyase – crystal structures of native phage–encoded hyaluronate lyase and its complexes with ascorbic acid and lactose docx
... length and 14 A in length between subsites and and and 3, respectively An examination of these regions indicates that PheC175 protrudes into the substrate binding area between subsites and Thus, ... and 320–334 Ascorbic acid and lactose molecule binds at three subsites in the polysaccharide binding site of the TSbH domain of HylP2 The positions of ligands are indicated at the substrate binding ... between sub˚ between subsites and 3, respecsites and and 14 A tively The first molecule of ascorbic acid is held at the lower-most subsite consisting of residues LysC179 and GluB167 LysC179 forms...
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Báo cáo khoa học: Using directed evolution to improve the solubility of the C-terminal domain of Escherichia coli aminopeptidase P Implications for metal binding and protein stability pptx
... diagram of the AMPP metal- binding sites Metal- binding ligands are Asp260, Asp271, His354, Glu383, and Glu406 (B) Stereo view of the AMPP active site Two mutations (Glu270 and Glu406) are responsible ... form ) is connected with its inability to bind metals Metalloproteins can fold via metal- dependent or metal- independent pathways [20,21] They may bind metal ions before polypeptide folding, after ... must be folded before metals bind ) the metal- binding ligands must be appropriately placed to coordinate the incoming metals Four acid residues coordinate the two divalent metal ions in the active...
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Báo cáo khóa học: Metal-binding stoichiometry and selectivity of the copper chaperone CopZ from Enterococcus hirae pot
... copz-ap6 ể FEBS 2004 EhCopZ metal binding and selectivity (Eur J Biochem 271) 995 Construction of the synthetic gene Ehcopz and expression vector PQE-cop Protein and metal quantication for titration ... respiration reveals metal- specic targets, interaction with an importer, and alternative sites for copper acquisition J Biol Chem 277, 54905497 ể FEBS 2004 EhCopZ metal binding and selectivity (Eur ... involved in the metal- binding site, it is crucial that the protein remains reduced throughout the experiment A control experiment was performed under ể FEBS 2004 EhCopZ metal binding and selectivity...
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Báo cáo Y học: Molecular interactions between nuclear factor kB (NF-kB) transcription factors and a PNA– DNA chimera mimicking NF-kB binding sites doc
... chimera mimicking the HIV-1 NF-kB binding sites Differential effects of the HIV-1 NF-kB PDP–PDP chimera on binding of NF-kB p52 and p50 to NF-kB binding sites of HIV-1 and IgK gene The experiment reported ... DNA and double stranded PDP –PDP chimeras mimicking the HIV-1 NF-kB binding sites, while effective inhibitors of binding of NF-kB p52 and p50 to HIV-1 LTR sequences, not efficiently inhibit the binding ... unlike symmetric NF-kB binding sites, possible problems related to self and/ or interstrand hybridization are expected to be minimal in the case of asymmetric NF-kB binding sites For these reasons,...
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Báo cáo khoa học: DG-based prediction and experimental confirmation of SYCRP1-binding sites on the Synechocystis genome pot
... putative binding sites relative to the ORF start position d A Sequences of putative binding sites e Changes in binding free energy and standard errors (DDGtotal Æ SE) Fig Confirmation of SYCRP1 binding ... al DG-based prediction of DNA binding sites by SYCRP1 Table Putative binding sites of SYCRP1 and the downstream genes The standard errors were calculated from standard errors of DDG No.a Locusb ... putative binding sites, we performed an EMSA to measure changes in binding free energy (observed DDGtotal) for the 11 binding sites with the lowest A DDGtotal values of the 23 putative binding sites...
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Báo cáo khoa học: Comparative metal binding and genomic analysis of the avian (chicken) and mammalian metallothionein potx
... metallothionein Results and discussion Zn(II) and Cd(II) binding abilities of ckMT and its separate domains Metal binding analysis rationale The metal binding abilities of ckMT were analysed following ... Fernando LP & Andrews GK (1989) Cloning and expression of an avian metallothionein-encoding gene Gene 81, 177–183 Fernando LP, Wei D & Andrews GK (1989) Structure and expression of chicken metallothionein ... ckMT, ackMT and bckMT metal complexes Spectroscopic (UV-Vis) and spectropolarimetric (CD) analysis of the metal ckMT clusters and of the species formed in vitro during the Zn ⁄ Cd and Zn ⁄ Cu...
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Báo cáo khoa học: An NMR study of the interaction between the human copper(I) chaperone and the second and fifth metal-binding domains of the Menkes protein pot
... Interaction between HAH1 and ATP7A does mutation of the metal- binding cysteines to serines [28] The data are thus consistent with a mechanism in which HAH1 and any of the ATP7A metal- binding domains interact ... putative intermolecular interaction region of yeast Atx1 and human HAH1 (upper), and of the various metal binding domains of yeast Ccc2 and human ATP7A (lower) Positively charged areas are blue, ... the surface of Atx1 and HAH1 is quite similar, but that of the metal binding domains of Ccc2 is somewhat different from ATP7A ⁄ ATP7B [11] In addition, although the two metal- binding domains of...
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Báo cáo khoa học: Aromatic amino-acid residues at the active and peripheral anionic sites control the binding of E2020 (AriceptÒ) to cholinesterases doc
... stabilize the binding of ligands such as huperzine A, edrophonium, acridines and one end of bisquaternary compounds such as BW284C51 and decamethonium [28,34,35] and destabilizes the binding of ... RTlnK¢I/KI, where K¢I and KI and are the dissociation constants for mutant and wild-type Mo AChE, respectively [28] Asp74 (Asp70 in BChE) are present in both AChE and BChE and Trp286 is replaced ... such as Asp74(72), Tyr72(70), Tyr124(121), and Trp286(279) contribute to the binding of E2020 to AChE Asp74 and Trp86 are present in both AChE and BChE, and the mutation of Trp86 (Trp82 in BChE)...
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Báo cáo Y học: Inhibition of SERCA Ca2+ pumps by 2-aminoethoxydiphenyl borate (2-APB) 2-APB reduces both Ca2+ binding and phosphoryl transfer from ATP, by interfering with the pathway leading to the Ca2+-binding sites ppt
... potential sites Figure 10 shows these binding sites in detail and highlights the amino acids Tyr837, Phe834, Phe256 and Asn768 One potential site is located between the top of transmembrane helix M7 and ... spectrophotometer and the absorbance was monitored at 422 and 390 nm, and the difference taken Ca2+ -binding to the ATPase was measured using the dual labeling technique of Longland et al [13] ATPase ... catalytic and regulatory sites were unchanged (i.e 9.4 ± 2.8 and 1.3 ± 0.5 mM, respectively) The Vmax values, however, were reduced The catalytic and regulatory Vmax values were 2.5 ± 0.1 and 5.4...
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