Identification of MMP Substrates in the Mammary Gland
Abstract
Matrix metalloproteinases (MMPs) are a family of zinc-dependent extracellular endopeptidases that have traditionally been viewed as effectors of late stages of cancer evolution. Recently, MMP-3 and MMP-14 were shown to act as natural tumor promoters when overproduced in mouse mammary glands (Sternlicht et al., 1999; Ha et al., 2001). To understand how these MMPs can alter cell behavior and influence breast cancer susceptibility, the yeast two-hybrid system was used to identify interacting proteins as potential novel MMP substrates. Three different bait' constructs were generated for each mouse and human MMP, but only the hemopexin domain hybrids appear suitable for screening. The catalytic domain hybrids are sticky' and auto-activate one or both reporters. The full-length hybrids also contain this sticky' catalytic domain and are probably poorly folded, as mature MMPs expressed in the yeast cytoplasm are inactive. Screening of a mouse and human mammary gland cDNA library form ATCC identified to date only intracellular interacting proteins. As the mRNA sources for these libraries were less than optimal, we may look for other libraries or make our own. In the latter case rough-ER bound mRNA will be isolated, to enrich for clones encoding membrane and secreted proteins.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2003
- Accession Number
- ADA423278
Entities
People
- Gerrit J. Dijkgraaf
Organizations
- University of California, San Francisco