Purification and characterization of two collagenase inhibitors from mouse sarcoma 180 conditioned medium

✍ Scribed by Rosalind A. Rosenthal; Marsha A. Moses; Yasushi Shintani; Joseph F. Megyesi; Robert Langer; Judah Folkman

• Book ID: 102877710
• Publisher: John Wiley and Sons
• Year: 1994
• Tongue: English
• Weight: 794 KB
• Volume: 56
• Category: Article
• ISSN: 0730-2312
• DOI: 10.1002/jcb.240560114

✦ Synopsis

We have previously shown that mouse sarcoma 180 cells produce vascular endothelial growth factor [VEGF; Rosenthal et al., 1990, Growth Factors, 4: 53-59], an endothelial mitogen that stimulates angiogenesis. Recent reports have implicated metalloproteinases and their inhibitors in the regulation of vascular morphogenesis, tumor invasion, and metastasis. We report here that mouse sarcoma 180 cells produce two collagenase inhibitors. These inhibitors were purified by heparin-Sepharose affinity chromatography, gel filtration, and C4 reverse phase h.p.1.c. Analytical gel electrophoresis of the purified inhibitors (MS-22 and MS-31) revealed molecular masses of 22,000 and 31,000 Da under reducing conditions, and 20,000 and 30,000 Da under nonreducing conditions, respectively. The NH,-terminal amino acid sequence of MS-22 was identical to that of tissue inhibitor of metalloproteinases type 2 (TIMP-2) produced by human melanoma cells [