An Improved Procedure for the Preparation of Thrombin Low Molecular Weight Substrates - Peptide p-Nitroanilides

Main Article Content

A.A Chistov
A.V. Talanova
M.V. Melnikova
S.S. Kuznetsova
E.F. Kolesanova

Abstract

Low molecular weight chromogenic thrombin peptide substrates, p-nitroanilides of short peptides protected at their N-terminal amino group, were prepared by solid-phase peptide synthesis on polystyrene-divinylbenzene polymer with trityl groups with preliminary attached p-phenylene diamine moiety. After the cleavage from the resin peptide p-aminoanilides were mildly oxidized to p-nitroanilides with the mixture of potassium sulfate and persulfate. Adsorption onto polymer support Bio-Beads SM-2 with further elution by acetonitrile allowed easy separating peptide p-nitroanilides from the oxidizer and obtaining the thrombin chromogenic substrate preparations with the target substance contents of not less than 95% and yields of 30-40%. Thrombin effectively catalyzed hydrolysis of the prepared substrates with KM and Vmax values of 29-134 mM and 0.03-1/16 mM/s, respectively.

Article Details

How to Cite
Chistov, A., Talanova, A., Melnikova, M., Kuznetsova, S., & Kolesanova, E. (2018). An Improved Procedure for the Preparation of Thrombin Low Molecular Weight Substrates - Peptide p-Nitroanilides. Biomedical Chemistry: Research and Methods, 1(4), e00057. https://doi.org/10.18097/BMCRM00057
Section
EXPERIMENTAL RESEARCH

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