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. 2012 May 10;287(27):22643–22653. doi: 10.1074/jbc.M111.337949

FIGURE 1.

FIGURE 1.

MMP-2 forms a reduction-sensitive homodimer. A, Western blot analysis using anti-Myc antibody on conditioned medium from COS-1 cells expressing pro-MMP-2-Myc/His under nonreducing and reducing conditions. β-ME, β-mercaptoethanol. Arrows indicate the monomeric and dimeric forms of pro-MMP-2 (n = 3). B, SDS-PAGE analysis of purified pro-MMP-2-Myc/His under nonreducing and reducing conditions. Pro-MMP-2-Myc/His was purified from the conditioned medium of the transfected cells using anti-c-Myc antibody-agarose. C, effect of SDS on dimer formation of MMP-2 in vitro. Purified pro-MMP-2-Myc/His (10 μg/ml) was treated with 10 mm iodoacetamide (IAA) at 37 °C for 1 h and then denatured with SDS-PAGE sample buffer in the presence of 10 mm iodoacetamide, followed by Western blot analysis using anti-Myc antibody (n = 2). IB, immunoblot. D, Western blot analysis using anti-Myc antibody of cell lysates from COS-1 cells expressing pro-MMP-2-Myc/His under nonreducing conditions. Cells were pretreated with 0.05% trypsin and 0.53 mm EDTA for 30 min on ice (n = 2).