Analysis of matrix metalloproteinase secretion by macrophages.
Matrix metalloproteinases (MMPs) are zinc-dependent proteases whose physiological roles include control of leukocyte migration. They are implicated in tissue destruction in inflammatory and infectious diseases. MMPs are not only capable of degrading all components of the extracellular matrix, but they also can modulate the immune response by cleaving cytokines and chemokines to alter their activity. Macrophages secrete a broad range of MMPs and represent a key source of MMPs in inflammatory lesions such as granulomas. Zymography is substrate-based gel electrophoresis that allows direct visualization of MMP activity. Here we describe measurement of MMP secretion from macrophages focusing on quantitative zymography. We also discuss complementary methods that should be used in parallel with zymography. The ability to analyze and quantify MMP secretion by macrophages offers an additional window through which to understand the contributions of macrophages to a wide variety of infectious, inflammatory, and immunologic disorders.
Department of Infectious Diseases & Immunity, Imperial College London, Hammersmith Campus, London, UK.
Paul T
PT
Elkington
Justin A
JA
Green
Jon S
JS
Friedland
D015153:Blotting, Western; D002364:Caseins / Q000378:metabolism; D002478:Cells, Cultured; D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000201:enzymology* / Q000378:metabolism*; D053505:Matrix Metalloproteinases, Secreted / Q000032:analysis*; D008863:Microspheres; D008967:Molecular Biology / Q000379:methods*; D011994:Recombinant Proteins / Q000032:analysis; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis
D002364:Caseins; D003470:Culture Media; D011994:Recombinant Proteins; D019715:Tissue Inhibitor of Metalloproteinase-1; D005780:Gelatin; D053505:Matrix Metalloproteinases, Secreted