PMID: 9428102Jan 15, 1998Paper

Thrombin-like enzyme from Lachesis muta muta venom: isolation and topographical analysis of its active site structure by means of the binding of amidines and guanidines as competitive inhibitors

Toxicon : Official Journal of the International Society on Toxinology
A MagalhãesE Rogana

Abstract

A serine protease enzyme was purified from Lachesis muta muta venom, with 40% yield, by gel filtration on Sephadex G-100 and affinity chromatography on Sepharose-agmatin. Homogeneity of the enzyme preparation was demonstrated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and the enzyme had a relative mol. wt of 45,000. The molar extinction coefficient at 280 nm was 62,127 (M x cm)-1. The enzyme hydrolysed Bz-Arg-Nan with Ks = 0.233 +/- 0.08 mM and kcat = 2.80 +/- 0.07 sec-1. All the amidines and guanidines tested for their inhibitory effect on thrombin-like enzyme behaved as competitive inhibitors of the enzyme with Ki values in the range 6.2 microM to 42.3 mM for amidines and 0.19 mM to 9.31 mM for guanidines. Dissociation constant values were analyzed in terms of the binding of the inhibitors with the subsite S1, the specificity pocket of the enzyme, Ki values were discussed in accordance with those for trypsin inhibition. beta-Naphthamidine was the strongest inhibitor, while guanidine was the weakest. The differences among the Ki values were interpreted in terms of the shape of the enzyme active site. For meta- and para-substituted benzamidinium ions a good correlation was found between log l/Ki and sigma Hamm...Continue Reading

References

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Citations

Oct 8, 2003·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Arinos MagalhaesEladio F Sanchez
Sep 12, 2012·Toxicon : Official Journal of the International Society on Toxinology·Cristiane Bregge-SilvaEliane Candiani Arantes
Nov 5, 2016·Toxicon : Official Journal of the International Society on Toxinology·Lourdes DiasStephen Hyslop
Feb 17, 2006·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·Arinos MagalhãesEladio F Sanchez
Apr 28, 2004·Bioorganic & Medicinal Chemistry Letters·Cyrille BoussardIan H Gilbert
Jul 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Salvatore Giovanni De-SimoneHerbert Leonel de Mattos Guedes
Oct 2, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Yu XinRongxiu Li

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