Matrix metalloproteinase (MMP)-2 and MMP-9 activities in human seminal plasma

Abstract

We report on the existence of two kinds of matrix metalloproteinases (MMPs), MMP-2 and MMP-9, in human seminal plasma. Partial purification of the proteinases was achieved by two steps, consisting of chromatography on a gel-filtration column and then on a gelatin affinity column. Proteinase activities in the chromatography extracts were shown to hydrolyse a fluorescent substrate specific to MMPs (Dnp-Pro-Leu-Gly-Leu-Trp-Ala-D-Arg-NH2). The proteinases were detected using gelatin-zymography, but were not detected using casein-zymography, and were also inhibited by EDTA, EGTA and o-phenanthroline. Molecular weights of the proteinases were determined by SDS-PAGE, gelatin-zymography and Western blot to be approximately 92, 84, 72, 67, 52 and 45 kDa. Gelatin-zymography showed three major bands of activity at 72, 67 and 52 kDa and minor bands at 92, 84 and 45 kDa. Apart from the two smallest bands, these proteinases were all recognized by the polyclonal antibodies for MMP-2 or MMP-9. These results indicate that two kinds of pro-form and active-form matrix metalloproteinases, MMP-2 and MMP-9, and their degradation products, are present in human seminal plasma.

Gelatinases in Boar Seminal Plasma and Their Relation to Semen Indicators

“…Based on previous reports (Metayer et al 2002;Shimokawa et al 2002;McCauley et al 2001;Tentes et al 2007), the bands with 225, 78 and 66 kDa correspond to dimer-MMP-9, proMMP-2 and active MMP-2. The variation in molecular weights (by 10 to 15 kDa) of gelatinases of different origins can be result of different glycosylation (Metayer et al 2002).…”

“…The variation in molecular weights (by 10 to 15 kDa) of gelatinases of different origins can be result of different glycosylation (Metayer et al 2002). Bands with lower molecular weights can represent the degradation products of metalloproteases with enzymatically active domain (Shimokawa et al 2002). This can be explained because both MMP-2 and MMP-9 have gelatin binding domains in their catalytic domains (Banyai et al 1994).…”

“…Metalloproteases, serine proteinases and serine proteinase inhibitors have been detected in turkey seminal plasma (Kotlowska et al 2005). Complexes of gelatinases and TIMP-1 and TIMP-2 (Shimokawa et al 2003) or MMP-2 and MMP-9 (Shimokawa et al 2002) were found in human seminal plasma. The first report of gelatinase in human sperms indicated higher 28-kDa gelatinase activity and lower 92-kDa MMP activity in normal compared to abnormal semen (Buchman-Shaked et al 2002).…”

The Impact of G1575A Matrix Metalloprotease-2 Gene Polymorphism on Male Fertility

“…Our results suggested that increasing enzymatic activity of MMP-2 might play a role in male infertility. Shimokawa et al showed that there is an active-form and a pro-form of Matrix Metalloproteinase-2 and its degradation products are present in human seminal plasma (12). In vitro studies have shown that the Follicle Stimulating Hormone (FSH) increases expression of MMP-2 in testicular sertoli cells (11).…”

“…Gelatinases are one of the subtypes of matrix metalloproteinases and includes matrix metalloproteinases 2 and 9 (collagenases A and B with molecular weight of 72 and 92 kD, respectively). These enzymes usually degrade gelatin and collagen type 4 (12)(13)(14). Matrix Metalloproteinase-2 is synthesized and secreted as a zymogen so the activation of MMP-2 is an important step in controlling its activity.…”