Assessment of gelatinases (MMP-2 and MMP-9) by gelatin zymography.
Gelatin zymography is a simple yet powerful method to detect proteolytic enzymes capable of degrading gelatin from various biological sources. It is particularly useful for the assessment of two key members of the matrix metalloproteinase family, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), due to their potent gelatin-degrading activity. This polyacrylamide gel electrophoresis-based method can provide a reliable assessment of the type of gelatinase, relative amount, and activation status (latent, compared with active enzyme forms) in cultured cells, tissues, and biological fluids. The method can be used to investigate factors that regulate gelatinase expression and modulate zymogen activation in experimental systems. The system provides information on the pattern of gelatinase expression and activation in human cancer tissues and how this relates to cancer progression. Interpretation of the data obtained in gelatin zymography requires a thorough understanding of the principles and pitfalls of the technique; this is particularly important when evaluating enzyme levels and the presence of active gelatinase species. If properly used, gelatin zymography is an excellent tool for the study of gelatinases in biological systems.
Department of Pathology, School of Medicine, Proteases and Cancer Program, Karmanos Cancer Institute, Wayne State University, Detroit, MI, USA.
Marta
M
Toth
Anjum
A
Sohail
Rafael
R
Fridman
D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D005780:Gelatin; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000302:isolation & purification / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000302:isolation & purification / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000737:chemistry
D011092:Polyethylene Glycols; D005780:Gelatin; D017830:Octoxynol; C010615:Nonidet P-40; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1