Expression and purification of membrane-type MMPs.
Matrix metalloproteinases (MMPs) modulate cell-cell and cell-matrix interactions and the membrane-type MMPs (MT-MMPs) are especially important for pericellular proteolysis. Understanding the activity, regulation, and function of MT-MMPs would provide important insight for ongoing research in many diseases including cancer, fibrosis, and autoimmune diseases. In this chapter, we introduce the methods to express and purify recombinant MT-MMPs using Escherichia coli or mammalian cells, as well as how to detect the activity of MT-MMPs in proMMP-2 activation using gelatin zymography.
Nie
Jing
J
Hope Heart Program, Benaroya Research Institute at Virginia Mason, Seattle, WA, USA.
Pei
Duanqing
D
D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism; D004926:Escherichia coli; D005780:Gelatin / Q000378:metabolism; D018093:Gelatinases / Q000378:metabolism; D006801:Humans; D053504:Matrix Metalloproteinases, Membrane-Associated / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*
D004792:Enzyme Precursors; D005780:Gelatin; D018093:Gelatinases; D053504:Matrix Metalloproteinases, Membrane-Associated; C069860:progelatinase