Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Synthesis of Carbobenzoxy-alanyl-thiaarginine (thialysine) benzyl ester and kinetic Studies with Trypsin

  • Published : 1998.06.20
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Abstract

Carbobenzoxy-alanyl-thiaarginine benzyl ester and carbobenzoxy-alanyl-thialysine benzyl ester were synthesized in solution. Kinetic studies were carried out using three different analytical methods, semi-classical method, progress curve analysis and competitive spectrophotometry. In competitive spectrophotometry, carbobenzoxy-valyl-glycyl-arginyl-p-nitroaniline was used as a detector. Kinetic constants such as $K_m$ and $V_{max}$ measured by competitive spectrophotometry are almost the same as those values measured by semi-classical method. Colorimetric Ellman's assays showed the thio-peptido mimetics to be a suitable substrates for trypsin. Kinetic studies with trypsin gave $K_m$ of 2.33 mM and $k_{cat}$ of $1.50{\times}10^5\;min^{-1}$ for carboxy-alanyl-thiaarginine benzyl ester and $K_m$ of $3.41{\times}10^{-3}\; Mm\; and\; k_{cat}\; of\; 520{\times}102\; min^{-1}$ for carbobenzoxy-alanyl-thialysine benzyl ester, respectively. Kinetic constants $(K_m=2.04{\times}10^{-2}\; mM, K_{cat}=4.42{\times}10^3 \;min^{-1})$ for natural substrate, carbobenzoxy-alanyl-lysine benzyl ester, were also evaluated by competitive spectrophotometry in order to compare the mode of binding on trypsin.

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