The Conformation of the Lysyl Side Chain of Substrates at the Active Center of Trypsin

• Published in: Journal of biochemistry (Tokyo) Vol. 100; no. 1; pp. 21 - 25
• Main Authors: MIZUSAKI, Koichi, SUGAHARA, Yuji, TSUNEMATSU, Hideaki, MAKISUMI, Satoru
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.07.1986
• Subjects: Analytical, structural and metabolic biochemistry; Binding Sites; Biological and medical sciences; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Hydrolases; Hydrolysis; Indicators and Reagents; Kinetics; Lysine - analogs & derivatives; Protein Conformation; Substrate Specificity; Trypsin - metabolism; Substrate; Trypsin; Side chain; Molecular structure; Lysine; Active site; Kinetic parameter; Substrate enzyme complex; Radiocrystallography; Conformation
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/oxfordjournals.jbchem.a121695

In order to study the conformation of the side chain of lysine substrates bound to the active center of trypsin, two lysine analogs, cis- and trans-2,6-diamino-4-hexenoic acids (4,5-dehydrolysines) were synthesized and kinetic parameters for the hydrolysis of benzoyl methyl esters and phenylthiazolones of these analogs by this enzyme were compared with those of the corresponding lysine derivatives. The derivatives of cis-4,5-dehydrolysine were hydrolyzed much more slowly than those of lysine, owing largely to the small kcat values for the former. On the other hand, the derivatives of the trans-isomer were hydrolyzed at about the same rates as those of lysine and the values of both Km and kcat of the former are also similar to those of the latter. These results indicate that the conformation of the side chain of the lysine derivatives hydrolyzed by trypsin is such that the β- and ε-carbons are in a trans-tike conformation, as suggested by X-ray crystallographic studies of inhibitor-trypsin complex.