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REINVESTIGATION OF THE REACTION OF CHYMOTRYPSIN WITH N-FURYLACRYLOYLTRYPTOPHAN DERIVATIVES AT ACIDIC PH

REINVESTIGATION OF THE REACTION OF CHYMOTRYPSIN WITH N-FURYLACRYLOYLTRYPTOPHAN DERIVATIVES AT ACIDIC PH BIOCHEMICAL JOURNAL Fink, A. L., Feldman, R., Zehnder, J. 1979; 181 (3): 733-736

Abstract

The reaction of alpha-chymotrypsin with N alpha-3-(2-furyl)acryloyl-L-tryptophan methyl ester (FA-Trp-OMe) and amide has been investigated in aqueous and dimethylsulphoxide cryosolvent solutions from pH2 to 7 and over a wide temperature range. Previous reports have suggested that an intermediate preceding the acyl-enzyme can be detected spectrophotometrically in the reaction with methyl esters of FA-Trp and FA-Tyr at low pH [Yu & Viswanatha (1969) Eur. J. Biochem. 11, 347--352), and that this intermediate is an oxazolinone [Coletti-Previero et al. (1970) FEBS Lett. 11, 213--217]. We show that the previous interpretations of the time-dependent spectral changes were incorrect, and that the only detected intermediate is the acyl-enzyme. This may be isolated by gel filtration at pH less than 2.5, 1 degree C, owing to its relative stability. The pH-dependence of the rates of acylation and deacylation from pH 8.5 to 2.0 are consistent with a single ionization of pK congruent to 7.0 in both aqueous and cryosolvent solutions.

View details for Web of Science ID A1979HL98500027

View details for PubMedID 42387

View details for PubMedCentralID PMC1161214