Quantitative Determination of High Molecular Weight Serum Proteinase Inhibitors in Human Semen

Abstract

Human seminal plasma contains high molecular weight and low molecular weight proteinase inhibitors. High molecular weight proteinase inhibitors are acid-lable and probably originate from serum. The following serum proteinase inhibitors are wellknown: alpha1 mtitrypsin, alpha1 ,x-antichymotrypsin, inter-alpha-trypsin inhibitor, antithrombin 111, alpha2-neuramino-glycoprotein and alpha2-macroglobulin (Heimburger et al. -1971).In contrast low molecular weight proteinase inhibitors are acidstable and are specific products of the male genital tract secretions (Fink et al. -1971;Zaneveld et al. -1971).The latter group consists of a trypsin-chymotrypsin inhibitor (HUSH, MW 11,000) and a trypsin-acrosin inhibitor MW 6,200). HUSI-I1 is a potent inhibitor of the sperm specific proteinase acrosin (Schiessler et al. -1975) involved in the penetration of the zona pellucida by the spermatozoa (Zaneveld et al. -1972).Since there are only few data available on high molecular weight serum proteinase inhibitors in human semen (Herrmann and Hermann -1969;Schumacher -1969) quantitative determinations of serum proteinase inhibitors were performed in seminal plasma of andrological patients using the radial immunodiffusion technique. Material and Methodsservice of the Department of Dermatology was collected immediately after masturbation (4-6 days of sexual abstinence). Medical history, clinical examination and semen evaluation showed no signs of chronic or acute genital tract inflammation. Semen analysis was performed according to the recommendations of the European Andrology Club. One hour after collection the seminal plasma was separated (first step: 600 x g, 15 min; second step: 16,000 x g, 5 min) and stored until used at -28OC in a deep freezer. Seminal vesicle fluids of two patients with carcinoma of the prostate (age: 52 and 54 years) were collected immediately after prostatectomy by puncturing the glands with a micropipette carefully avoiding blood contamination.Antisera from rabbit containing monospecific antibodies against the following human serum proteinase inhibitors were used: Alpha1-antitrypsin (Behring Werke AG, Batch No. 2296 AA), alphai,+ntichymotrypsin (Behring Werke AG, Batch No. 1926 BL), inter-alpha-trypsin inhibitor (Behring Werke AG, Batch No. 1916 Z), antithrombin I11 (Behring Werke AG, Batch No. 2252 D), alpha2-neuramino-glycoprotein (Behring Werke AG, Batch No. 1807 BE) and alpha2-macroglobulin (Behring Werke AG, Batch No. 151 1 AN). Protein standards for the different inhibitors were obtained from Beh-Material: The semen of 129 male subjects (age: 23-53 years) attending the infertility

Alpha-1 antitrypsin phenotypes in patients with Klinefelter’s syndrome

“…Our finding coincided with the data, which have shown that azoospermic ejaculates had lower mean AAT concentration compared to oligo-zoospermic and normo-zoospermic ejaculates (Schill 1976). Handelsman et al (1986) found reduced semen volume in infertile men with homozygous ZZ AAT deficiency.…”

Possible effects of the kallikrein-kinin system on male reproductive functions

“…The main kallikrein inhibitor in the seminal plasma is a, proteinase inhibitor which comes from the serum where its level is 20 times higher than in the seminal plasma (Schill, 1976). However, they are rapidly inhibited by low molecular weight proteinase inhibitors found in seminal fluids.…”

“…This raises the question of which kininogenase inhibitors are present in male genital tract secretions. The main kallikrein inhibitor in the seminal plasma is a, proteinase inhibitor which comes from the serum where its level is 20 times higher than in the seminal plasma (Schill, 1976). a, proteinase inhibitor is a progressive type of kallikrein inhibitor which does not lead to an immediate inactivation of tissue kallikrein in seminal plasma.…”

Complexes of Gelatinases and Tissue Inhibitor of Metalloproteinases in Human Seminal Plasma

“…H uman seminal plasma contains several kinds of inhibitor that are secreted from the prostate gland. The existence of many proteinase inhibitors that contain trypsin-chymotrypsin inhibitor (HUSI-I) and trypsin-acrosin inhibitor (HUSI-II), cysteine proteinase inhibitors (In-A and In-B), high-molecular-weight serum proteinase inhibitors, protein C inhibitor and acrosome reaction-inhibiting glycoprotein in human seminal plasma has been reported (Schiessler et al, 1976;Schill, 1976;Minakata et al, 1986;Christensson and Lilja, 1994;Drisdel et al, 1995). However, the existence of TIMPs that contain complexes of gelatinases and TIMPs is relatively unknown in human seminal plasma.…”