Translator Disclaimer
1 April 2004 Distinction of Sperm-Binding Site and Reactive Site for Trypsin Inhibition on P12 Secreted from the Accessory Sex Glands of Male Mice
Author Affiliations +
Abstract

Six variants of P12, a Kazal-type trypsin inhibitor in the secretion of male mouse accessory sexual glands, were made using single-site mutations including R19L, Y21V, D22G, R43G, K44S, and R45T, based on one-letter-code mutation of amino acids. The other two variants, Nd10 and Cd8, were made using the deletion of 10 and 8 residues from the N- and C-terminals, respectively. Their CD profiles revealed maintenance of the P12 conformation in the seven variants, excluding Cd8, which became unfolded. Only R19L entirely lost the ability while the other variants were as strong as P12 in inhibiting the trypsin digestion of N-benzoyl-Phe-Val-Arg 7-amido-4-methylcoumarin. The immunocytochemical results demonstrated that D22G and Cd8 failed to bind to sperm, Y21V very weakly did so, and the other variants retained their sperm-binding abilities. Concomitantly, the immunocytochemical stainability of each ligand was parallel to its inhibitory effect on 125I-P12-sperm binding, and a synthetic oligopeptide corresponding to residues 18–24 of P12 was able to inhibit P12-sperm binding. The data together concluded that R19 was essential for protease inhibition and D22 and/or Y21 mainly being responsible for the binding of P12 to sperm. The steric arrangement of R19, Y21, and D22 on the tertiary structure of P12 is discussed.

Ching-Wei Luo, Han-Jia Lin, S. C. B. Gopinath, and Yee-Hsiung Chen "Distinction of Sperm-Binding Site and Reactive Site for Trypsin Inhibition on P12 Secreted from the Accessory Sex Glands of Male Mice," Biology of Reproduction 70(4), 965-971, (1 April 2004). https://doi.org/10.1095/biolreprod.103.020552
Received: 23 June 2003; Accepted: 1 November 2003; Published: 1 April 2004
JOURNAL ARTICLE
7 PAGES

This article is only available to subscribers.
It is not available for individual sale.
+ SAVE TO MY LIBRARY

SHARE
ARTICLE IMPACT
RIGHTS & PERMISSIONS
Get copyright permission
Back to Top