`Collagenase-Like' Peptidase in Human, Rat and Bull Spermatozoa

Abstract

Summary. A peptidase capable of splitting synthetic collagenase substrate and collagen peptides,`collagenase-like' peptidase, has been discovered in human semen, rat and bull epididymal spermatozoa and rat and bull testes. This enzyme is also present in human seminal plasma and sperm washings, but its total activity pattern in human semen fractions, its specific activity in washed spermatozoa and the negligible activity found in the azo\l=o"\spermicejaculate prove that it is a sperm enzyme. As shown by the thin-layer chromatography of the reaction products,`collagenase-like' peptidase cleaves synthetic collagenase substrate at the same site as bacterial collagenase does. The pH optimum of the`c ollagenase-like' peptidase is 7\m=.\5.The enzyme has been partially purified from rat testis homogenate by ammonium sulphate saturation and gel filtration on a Sephadex G-200 column. Its molecular weight appears to be about 110,000. The possible r\l=o^\leof this enzyme in fertilization is discussed.

The metalloenzymic nature of collagenase-like peptidase of the rat testis

“…Although CLP can not be considered a 'true collagenase' (it does not degrade native collagen), it is obviously similar to collagenases, not only because it is capable of degrading a synthetic collagenase substrate and collagen peptides (Koren & Milkovic, 1973), but also in respect to its metal requirement. The fact that zinc is one of the metals inhibiting CLP activity (Table 1) does not exclude the possibility that this metal is present at the active site of the enzyme, since it is known that a metal present at the active centre of an enzyme molecule may exert an inhibitory effect when it is also bound to some other part of the same molecule (Vallee, 1960).…”

“…We have previously described a collagenase-like peptidase (CLP), an enzyme capable of degrading synthetic collagenase substrate, whose activity was detected in rat and bull testes and spermatozoa and in human spermatozoa (Koren & Milkovic, 1973). While investigating the role of CLP in the male reproductive system we found that purified CLP from rat testes degraded proteins from the rat seminal vesicle secretion, and could thus promote the release of spermatozoa from the copulatory plug (Koren, Lukac & Milkovic, 1974;Koren, Schön & Lukac, 1975).…”

Feasibility of sex-sorting sperm from the white and the black rhinoceros (Ceratotherium simum, Diceros bicornis)

“…In the human, a chymotrypsin-like prostatic-specific antigen was determined to cause liquefaction of semen through cleavage of semenogelin I, the major component of the coagulum, which is expressed exclusively in the vesicular glands [23][24][25]. Additionally, proteolytic enzymes were detected and isolated in the seminal plasma of different species such as collagenase-like metalloproteinases in human, rat, and bull semen [26] and trypsin in semen from the guinea pig and rabbit [27,28]. Based on these findings, several enzymes have been used to cleave the gelatinous components of seminal plasma in order to reduce its viscosity.…”

The Coagulation of Insoluble and Basic Proteins From Rat Seminal Vesicle Secretion With Vesiculase: Influence of Collagenaselike Peptidase From Rat Testis

“…The effects of collagenase-like peptidase (CLP) (Koren & Milkovic, 1973;Koren, Lukac & Milkovic, 1974) on seminal vesicle secretion fractions indispensable for formation of the copulatory plug are presented and a rôle for sperm CLP in the release of spermatozoa from the plug is suggested.…”