Detection of Matrix Metalloproteinases by Zymography

Abstract

Matrix metalloproteinases (MMPs) represent more than 20 zinc-containing endopeptidases that cleave internal peptide bonds, leading to protein degradation. They play a critical role in many physiological cell functions, including tissue remodeling, embryogenesis, and angiogenesis. They are also involved in the pathogenesis of a vast array of diseases, including but not limited to systemic inflammation, various cancers, and cardiovascular, neurological, and autoimmune diseases. Here, we describe gel zymography to detect MMPs in cell and tissue samples and in cell culture supernatants.

Keywords: Detection; Protease; Semiquantitative; Zymogram; Zymography.

Figure 1

Gelatin zymography gel loaded with different dilutions of horse serum (HS), goat serum (GS), and human serum (HuS) (1, 5, 10%). Stronger bands of MMP-2 (~ 65 kD) and MMP-9 (~ 85 kD) are visible with increasing concentrations of the different sera.

Figure 2

Gelatin zymography gel comparing MMP-2 and MMP-9 production in the supernatant of untreated fibroblast-like synoviocytes isolated from a patient with rheumatoid arthritis (RA-FLS; control) and paxilline-treated RA-FLS incubated in the presence of 0, 1, 5, or 10% of fetal bovine serum. Paxilline-treated RA-FLS exhibit a decrease in MMP-2 production when incubated without serum. This effect is masked by incubation with 1, 5, 10% serum.