Xenbase Image ID: 134369

Fig. 1. Gelatin zymography and Western blot of the extract obtained from mature sperm after SDS-PAGE with 10% gels. (A) Gelatin zymography of the sperm extract showing major 68 kDa and 55 kDa lytic bands and a minor 60 kDa band (lane 1), but no lytic band in the presence of 1 mM o-phenanthroline during gel-incubation (lane 2). (B) Western blot of the sperm extract by anti-MMP-2 catalytic antibody (lane 1), showing a major band of 68 kDa band and minor 55, 60 and 53 kDa bands. Western blots of the sperm extract by anti-MMP-2 HPX antibody, showing 53-28 kDa bands lower than 68-55 kDa bands (lane 2). Western blot of the sperm membrane fraction (lane 3) and the sperm cytoplasmic fraction (lane 4) by anti-MMP-2 HPX antibody, showing 68-55 kDa bands and several minor bands with the HPX domain (53-28 kDa) in the membrane fraction, but a major 55 kDa band and minor 42 and 39 kDa bands in the cytoplasmic fraction. (C) Immunoprecipitation of the sperm extract by anti-MMP-2 HPX antibody, showing precipitation of MMP-2 (68-53 kDa) (lane 1) and coprecipitation of a 106-76 kDa fraction of SGP (lane 3), but no precipitation by pre-immune antibody (lanes 2 and 4). Asterisks, input antibodies. (D) A domain structure of MMP-2, showing the signal peptide (Pre), the prodomain (Pro), the catalytic domain (Cat), the fibronectin-like collagen-binding domain (FN), the zinc-binding site (Zn2+), and the hemopexin domain (HPX). Image published in: Iwao Y et al. (2014) Copyright © 2014. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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