Human a-thrombin

Bode, W., et al. The refined 1.9 A crystal structure of human a-thrombin interaction with D-Phe-Pro-Arg chloromethylketone and significance of the Tyr-Pro-Pro-Trp insertion segment. EMBO ]. 8 3467-3475,... 

Kubik M. F Stephens A. W., Schneider D et al. High-affinity RNA ligands to human a thrombin. Nucleic Acids Res 1994 22,2619-26. 

Caflisch A. Computational combinatorial ligand design Application to human a-thrombin. J Computer-Aided Molec Design 1996 10 372-396. 

The effects of the mutations on functional responses were determined on thrombin receptors that were expressed in Xenopus oocytes. Agonist-stimulated calcium efflux with human a-thrombin (10 nM) and SFLLRNP-NH2 (40 pM), as described previously,29 was employed to determine the functional responsiveness of the receptor. 

Perdue JR Lubenskyi W, Kivity E, Sonder SA, Fenton JW, Protease mitogenic response of chick embryo fibroblasts and receptor binding/processing of human a-thrombin. J Biol Chem 1981 256 2767-2776. 

Fig. 3.10 Result of the limit of detection (LOD) determination for NS-TBA in binding buffer for the cognate target protein human a thrombin. The sensor was incubated with protein concentrations between 500 yM and 50 pM. The red line indicates the LOD (mean plus 3 standard deviations of the blank measurement) [49]...

Human a-thrombin and the fluorogenic substrate (Tos-Gly-Pro-Arg-AMC HCI) were purchased from Sigma. Fmoc derivatives of amino acids were purchased from Advanced ChemTech and Novabiochem. N-a-Fmoc-N-y-trityl-L-Gln-Wang resin was purchased from Applied Biosystem Inc. The solvents used in peptide synthesis were obtained from Anachemia Chemical Inc. and Applied Biosystems Inc. 

Schaeffer R Jr, Bitrick M Jr. Effects of human a-thrombin and 8-bromo-cAMP on large and microvessel endothelial monolayer equivalent pore radii. Microvasc Res 1995 49 364. 

Inhibitory dissociation constants for human a-thrombin mediated hydrolysis of the... 

Chirgadze, N.Y., Sail, D.J., Briggs, S.L., Clawson, D.K., Zhang, M., et al. (2000) The crystal structures of human a-thrombin complexed with active site-directed diamino benzo[b]thiophene derivatives a binding mode for a stmeturaUy novel class of inhibitors. Protein Sci. 9 29-36. 

Berliner L J, Shen Y Y L (1977). Physical evidence for an apolar binding site near the catalytic centre of human a-thrombin. Biochem. 16 4622-4626. 

Figure 2 Elution of human a-thrombin at 25°C on PAOM (A) and PSSO (B) resins with a linear gradient from 0.1 to 2 M sodium chloride in 0.05 M phosphate buffer, pH = 7.4 flow-rate, 1 ml/min.

SP Sephadex as before ( 1 0) using a modification of the method of Lundblad et al. (JJ ) to produce a product with a specific activity of 1850 NIH U/mg (). Purified human a-thrombin (3000 - 31 53 NIH U/mg) was the kind gift of Dr. J.W. Fenton II (N,Y. State Department of Health). Heat defibrinated citrated human plasma (5 °C, 5 min) was used as a source of crude antithrombin III (0.2 mg/mL). Purified human antithrombin III (1000 U/mg 1 mL plasma = 100 U) was the kind gift of Dr. M. Wickerhauser (American Red Cross, Bethesda, MD). A purified human antithrombin III of similiar activity to that of Wickerhauser (0.29 mg/mL) was also prepared from citrated human plasma (Red Cross Blood Bank, Toronto) using heparin-Sepharose (13). 

Heparin with high anticoagulant activity has been recovered from crude commercial preparations by affinity chromatography on columns of immobilized human a-thrombin. ... 

The covalent modification of human a-thrombin with pyridoxal 5 -phosphate involves phosphopyridoxylation at two sites.Attachment of the cofactor to the first site was accompanied by a loss in the fibrinogen clotting activity of the thrombin, whereas attachment of the cofactor to the second site resulted in a decrease in the sensitivity to heparin in the anti-(thrombin III)-thrombin reaction. 

Scheme 22.7 Schematic description of the specific recognition of human a-thrombin using single-stranded DNA thrombin aptamer and cationic polythiophene...

Figure 22.7 Specificity of the detection of human a-thrombin. Fluorescence intensity was measured at 570 nm with excitation at 408 nm. From M. Bera-Aberem, Najari, H. Ho, J. F. Gravel, P. Nobert, D. Boudreau and M. Leclerc, Protein detecting arrays based on cationic polythiophene-DNA-aptamer complexes, Adv. Mater., 18, 2703-2707 (2006). Copyright Wiley-VCH Verlag GmbH Co. KGaA. Reproduced with permission...

Design of smart MRI contrast agent based on SPIONs and aptamers is described for the detection of human a-thrombin protein... 

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