Open AccessIdentification of complexes of gelatinase A and tissue inhibitor of metalloproteinase‐2 in human follicular fluid
Author(s) -
Shimokawa KenIchi,
Katayama Masatoki,
Matsuda Yoshifumi,
Takahashi Hidenobu,
Hara Izumi,
Sato Hirohisa
Publication year - 2003
Publication title -
reproductive medicine and biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.005
H-Index - 22
eISSN - 1447-0578
pISSN - 1445-5781
DOI - 10.1046/j.1445-5781.2003.00029.x
Subject(s) - gelatinase , matrix metalloproteinase , tissue inhibitor of metalloproteinase , zymography , western blot , gelatinase a , metalloproteinase , chemistry , gelatin , follicular fluid , extracellular matrix , microbiology and biotechnology , follicular phase , gelatinases , blot , basement membrane , biochemistry , biology , endocrinology , embryo , oocyte , gene
Background and Aims: Ovulation involves considerable tissue remodeling in normal ovarian function. These processes are expected to involve matrix metalloproteinases (MMP). Follicular rupture is caused by the degradation of the basement membrane between the thecal and granulose layers, as well as disruption of the extracellular matrix (ECM) at the site of rupture. We report on the existence of the complexes of progelatinase A (proMMP‐2), MMP‐2 and a tissue inhibitor of metalloproteinase‐2 (TIMP‐2) using zymographic and immunological techniques in human follicular fluid (HFF). Methods and Results: Partial purification of the complexes was achieved by using gelatin affinity column chromatography. The peak (tubes 68–73) in this chromatography showed gelatinase activities by gelatin‐zymography, and also an inhibition by EDTA (metalloproteinase inhibitor). The molecular weights of the gelatinase activities were approximately 72 and 67 kDa, and were consistent with standard proMMP‐2 and MMP‐2, as found by using gelatin‐zymography. Similarly, the band in this peak was consistent with standard recombinant full‐length TIMP‐2, as found by the use of western blot analysis, and the molecular weight of the band was approximately 21 kDa. Conclusion: As proMMP‐2, MMP‐2 and TIMP‐2 exist in the peak from the gelatin affinity column, we expected that these form the complexes. These results indicate that the complexes of proMMP‐2/TIMP‐2 and MMP‐2/TIMP‐2 exist in HFF. (Reprod Med Biol 2003; 2 : 115–119)