Binding constants

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  • Each generation has its unique needs and aspirations. When Charles Wiley first opened his small printing shop in lower Manhattan in 1807, it was a generation of boundless potential searching for an identity. And we were there, helping to define a new American literary tradition. Over half a century later, in the midst of the Second Industrial Revolution, it was a generation focused on building the future. Once again, we were there, supplying the critical scientific, technical, and engineering knowledge that helped frame the world.

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  • Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học 'Respiratory Research cung cấp cho các bạn kiến thức về ngành y đề tài: " Open Access SP-A binds alpha1-antitrypsin in vitro and reduces the association rate constant for neutrophil elastase...

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  • The guanine nucleotide-binding protein Ras occurs in solution in two different conformational states, state 1 and state 2 with an equilibrium constant K12 of 2.0, when the GTP analogue guanosine-5¢-(b,c-imido)tri-phosphate or guanosine-5¢-(b,c-methyleno)triphosphate is bound to the active centre.

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  • Thein vivo ligand-binding function and ligand-binding activity of the Dro-sophila melanogaster retinoid-X receptor (RXR) ortholog, ultraspiracle, toward natural farnesoid products of the ring gland were assessed. Using an equilibrium fluorescence-binding assay, farnesoid products in the juven-ile hormone (JH) biosynthesis pathway, and their epoxy derivatives, were measured for their affinity constant for ultraspiracle (USP).

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  • Genistein and daidzein, the major isoflavones present in soybeans, possess a wide spectrum of physiological and pharmacological functions. The bind-ing of genistein to human serum albumin (HSA) has been investigated by equilibrium dialysis, fluorescence measurements, CD and molecular visuali-zation. One mole of genistein is bound per mole of HSA with a binding constant of 1.5 ± 0.2·10 5 m )1 . Binding of genistein to HSA precludes the attachment of daidzein.

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  • Observations of thioredoxin inhibition by cadmium and of a positive role for thioredoxin in protection from Cd 2+ led us to investigate the thioredoxin–cadmium interaction properties.We used calorimetric and spectroscopicmethods at different pHvalues to explore the relative contribution of putative binding residues (Cys32, Cys35, Trp28, Trp31 and Asp26) within or near the active site. At pH 8 or 7.5 two binding sites were identified by isothermal titration calorimetry with affinity constants of 10·10 6 M )1 and 1·10 6 M )1 . ...

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  • In the search for new ligands for the plasminogen kringle 4 binding-protein tetranectin, it has been found by ligand blot analysis andELISAthat tetranectinspecificallybound to the plasminogen-like hepatocyte growth factor and tissue-type plasminogen activator.The dissociation constants of these complexes were found to be within the same order of mag-nitude as the one for the plasminogen-tetranectin complex. The study also revealed that tetranectin did not interact with the kindred proteins: macrophage-stimulating protein, urokinase-type plasminogen activator and prothrombin....

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  • A comparative study of thermodynamic and kinetic aspects of Cu(II) and Ni(II) binding at the N-terminal binding site of human and bovine serum albumins (HSA and BSA, respectively) and short peptide analogues was performed using potentiometry and spectroscopic techniques. It was found that while qualitative aspects of interaction (spectra and structures of complexes, order of reactions) could be reproduced, the quantitative parameters (stability and rate constants) could not.

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  • The high-pressure stopped-flow technique is applied to study the CO binding in cytochrome P450cam (P450cam) bound with homologous substrates (1R-camphor, camphane, norcamphor and norbornane) and in the substrate-free protein. The activation volume DV # of the CO on-rate is positive for P450cam bound with substrates that do not contain methyl groups. The kon rate constant for these substrate complexes is in the order of 3 · 106 M)1Æs)1.

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  • Microfibrils and elastin are major constituents of elastic fibers, the assembly of which is dictated by multimolecular interactions.Microfibril-associated glycoprotein-1 (MAGP-1) is a microfibrillar component that interacts with the sol-uble elastin precursor, tropoelastin. We describe here the adaptation of a solid-phase binding assay that defines the effect of divalent cations on the interactions between MAGP-1 and tropoelastin. Using this assay, a strong cal-cium-dependent interaction was demonstrated, with a dis-sociation constant of 2.8 ± 0.

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  • The ability ofEntamoeba histolyticaTATA binding protein (EhTBP) to interact with different TATA boxes in gene promoters may be one of the key factors to perform an efficient transcription in this human parasite. In this paper we used several TATA variants to study the in vitro EhTBP DNA-binding activity and to determine the TATA-EhTBP dissociation constants. The presence of EhTBP in complexes formed by nuclear extracts (NE) and the TATTTAAA oligonucleotide, which corresponds to the canonical TATA box for E. histolytica, was demonstrated by gel-shift assays. ...

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  • High-resolution X-ray diffraction structures of integral membrane proteins have revealed various binding modes of lipids, but current spectroscopic studies still use uniform macroscopic binding constants to describe lipid binding. The Adair approach employing microscopic lipid-binding con-stants has previously been taken to explain the enhancement of agonist binding to the nicotinic acetylcholine receptor by general anaesthetics in terms of the competitive displacement of essential lipid activator molecules [Walcher S, Altschuh J & Sandermann H (2001) J. Biol. Chem. 276, 42191–42195]....

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  • S100A1 is a typical representative of a group of EF-hand calcium-binding proteins known as the S100 family. The protein is composed of twoasub-units, each containing two calcium-binding loops (N and C). At physiologi-cal pH (7.2) and NaCl concentration (100 mm), we determined the microscopic binding constants of calcium to S100A1 by analysing the Ca 2+ -titration curves of Trp90 fluorescence for both the native protein and its Glu32fiGln mutant with an inactive N-loop.

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  • Polyphenol oxidase (PPO) catalyzes the oxidation ofo-diphenols to their respective quinones. The quinones autopolymerize to form dark pigments, an undesired effect. PPO is therefore the target for the development of antibrowning and antimelanization agents. A series of phenolic compounds experimentally evaluated for their binding affinity and inhibition constants were computationally docked to the active site of catechol oxidase.

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  • Prostacyclin synthase (PGIS) is a member of the cytochrome P450 family in which the oxyferrous complexes are generally labile in the absence of substrate. At 4C, the on-rate constants and off-rate constants of oxygen binding to PGIS in solution are 5.9·10 5 m )1 Æs )1 and 29 s )1 , respectively.

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  • The different effects of pH and NaCl on individual O2-binding properties ofaandbsubunits within liganded tetramer and dimer of human hemo-globin (HbA) were examined in a number of laser time-resolved spectro-scopic measurements. A previously proposed approach [Dzhagarov BM & Lepeshkevich SV (2004)Chem Phys Lett390, 59–64] was used to determine the extent of subunit dissociation rate constant difference and subunit affinity difference from a single flash photolysis experiment.

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  • Thermobifida fusca xyloglucan-specific endo-b-1,4-gluca-nase (Xeg)74 and the Xeg74 catalytic domain (CD) were cloned, expressed inEscherichia coli, purified and charac-terized.This enzymehas aglycohydrolase family-74CDthat is a specific xyloglucanase followed by a family-2 carbo-hydrate binding module at the C terminus.The Michaelis constant (Km) andmaximal rate (Vmax) values for hydrolysis of tamarind seed xyloglucan (tamXG) are 2.4lMand 966lmol xyloglucan oligosaccharides (XGOs) min )1 Ælmol protein )1 ....

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  • The phosphoproteins HPrSerP and CrhP are the main effectors for CcpA-mediated carbon catabolite regulation (CCR) inBacillus subtilis. Complexes of CcpA with HPrSerP or CrhP regulate genes by binding to the catabolite responsive elements (cre). We present a quantitative analysis of HPrSerP and CrhP interaction with CcpA by surface plasmon resonance (SPR) revealing small and similar equilibrium constants of 4.8 ± 0.4 lm for HPrSerP–CcpA and 19.1 ± 2.5 lmfor CrhP–CcpA complex dissoci-ation.

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  • In aqueous media, muscle pyruvate kinase is highly selective for K + over Na + . We now studied the selec-tivity of pyruvate kinase in water/dimethylsulfoxide mix-tures by measuring the activation and inhibition constants of K + and Na + , i.e. their binding to the monovalent and divalent cation binding sites of pyruvate kinase, respect-ively [Melchoir J.B. (1965) Biochemistry4, 1518–1525]. In 40% dimethylsulfoxide theK0.5 appfor K + and Na + were 190 and 64-fold lower than in water.Kiappfor K + and Na +decreased 116 and 135-fold between 20 and 40% dimethylsulfoxide.

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  • The new antigen receptor (IgNAR) from sharks is a disul-phide bonded dimer of two protein chains,each containing one variable and five constant domains,and functions as an antibody. In order to assess the antigen-binding capabilities of isolated IgNAR variable domains (VNAR),we have con-structed anin vitro library incorporating synthetic CDR3 regions of 15–18 residues in length.

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