Date of Award
2009
Document Type
Thesis
Publication Status
Version of Record
Submission Date
October 2026
Abstract
The tissue inhibitors of metalloproteinases (TIMPs) are endogenous inhibitors of the matrix metalloproteinases (MMPs). Since unregulated MMP activities are linked to arthritis, cancer, and atherosclerosis, TIMP variants that are selective inhibitors of disease-related MMPs have potential therapeutic value. The structures of TIMP/MMP complexes reveal that most interactions with the MMP involve the N-terminal region of TIMP and the C-D B-strand connector which occupy the primed (right side of the active site) and unprimed (left side) regions of the active site. Substitutions for Thr2 of N-TIMP- 1 strongly influence MMP selectivity. In this study we found that Arg and Gly, which generally reduce MMP affinity, have less effect on binding to MMP-9. When the Arg mutation is added to the NTIMP-1 mutant with AB loop of TIMP-2, it produced a gelatinase-specific inhibitor with Ki values of 2.8 and 0.4 nM for MMP-2 and MMP-9, respectively. The Gly mutant has a Ki of 2.1 nM for MMP-9 and > 40 uM for MMP-2, indicating that engineered TIMPs can discriminate between MMPs in the same subfamily. In collaboration with Dr. Yingnan Zhang at Genentech, we have developed a protocol for the phage display of full-length human TIMP-2 to identify high-affinity selective inhibitors of human MMP-1, a protease that plays a role in cleaving extracellular matrix (ECM) components, connective tissue remodeling during development, angiogenesis, and apoptosis. We have generated a library containing 2x1010 variants of TIMP-2 randomized at residues 2-6 (L1), at residues 34-40 (L2) and 67-70 (L3).
Extent
xvii, 157 p. : ill. (some col.)
Recommended Citation
Bahudhanapati, Harinathachari., "In search of MMP specific inhibitors" (2009). Electronic Theses and Dissertations 2000 - 2009. 1702.
https://digitalcommons.fau.edu/etd_general_5/1702
Identifier
FADT221942
Comments
by Harinathachari Bahudhanapati.|Thesis (Ph.D.)--Florida Atlantic University, 2009.|Includes bibliography.|Electronic reproduction. Boca Raton, Fla., 2009. Mode of access: World Wide Web.|by Harinathachari Bahudhanapati.