PMID: 1204614Nov 1, 1975Paper

The reactions of alpha-chymotrypsin and related proteins with ester substrates in non-aqueous solvents

European Journal of Biochemistry
A A. KlyosovI V. Berezin

Abstract

1. The reactivity of alpha-chymotrypsin toward p-nitrophenylacetate has been studied in dimethylformamide, dimethylsulfoxide, formamide and methylacetamide. p-Nitrophenol is liberated in dimethylsulfoxide only. 2. The reactions of alpha-chymotrypsin in dimethylsulfoxide are characterized by the same kinetic and equilibrium constants with either the p-nitrophenyl esters of straight chain carboxylic acids (from acetic to n-caprylic) or with the "specific substrate", N-carbobenzoxy-DL-phenylalanine p-nitrophenyl ester. This signifies that reactions of alpha-chymotrypsin in dimethylsulfoxide, unlike those in aqueous medium, have no specificity toward su-strate structure. 3. The stoichiometry of alpha-chymotrypsin reactions in dimethylsulfoxide was shown to be about five moles of substrate per mole of enzyme. After attaining this stoichiometry, the reaction is completed. 4. Optical rotatory dispersion spectra indicate that in non-aqueous media alpha-chymotrypsin undergoes a large conformational transition which results in a random coil. 5. Chymotrypsinogen, trypsin, trysinogen, lysozyme and serum albumin react with p-nitrophenylacetate in dimethylsulfoxide at rates which are approximately equal to those of alpha-chymotrypsin. Thus, ...Continue Reading

References

May 11, 1973·Molecular and Cellular Biochemistry·P Douzou
Oct 30, 1974·Journal of the American Chemical Society·L M HinmanD A Deranleau
Jan 7, 1969·Biochimica Et Biophysica Acta·F SeydouxG Némethy
Jan 20, 1959·The Journal of General Physiology·M CASTANEDA-AGULLO, L M DEL CASTILLO

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Citations

Sep 1, 1977·Biotechnology and Bioengineering·A KlibanovI V Berezin
Mar 5, 1992·Biotechnology and Bioengineering·N Chang, A M Klibanov
Jun 5, 1994·Biotechnology and Bioengineering·J T ChinA M Klibanov
Jun 11, 2011·Biotechnology Letters·Takashi Kobayashi
Dec 1, 1976·Biochimica Et Biophysica Acta·V S Sethi, P Okano
Oct 1, 1992·Enzyme and Microbial Technology·T NagashimaH Kise
Mar 10, 1992·Biochemistry·K Ryu, J S Dordick
Jul 7, 1978·Biochimica Et Biophysica Acta·R K TripathiR D O'Brien
May 15, 1991·Biochemical and Biophysical Research Communications·N ChangA M Klibanov

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