The applicant, Bjorn Steffensen, DDS, MS, PhD, is applying for the Individual Scientist Award (K02) to support his research career development. The goals of this career development program are to: 1) Extend the ongoing research to identify approaches to inhibit enzymes involved in periodontal disease and oral cancer, 2) Gain new knowledge and skills on novel proteomics technologies to strengthen ongoing and future research, and 3) Evolve into a productive and independent scientist with a dynamic and progressive research program focused on the mechanisms of wound healing, periodontal disease, and oral cancer. The research plan addresses the mechanism by which matrix metalloproteinase-2 (MMP-2) degrades tissues. The MMP-2 belongs to the matrix metalloproteinase family of enzymes, which collectively can degrade most tissue components. The MMPs are beneficial features of normal development and tissue adaptation, but uncontrolled MMP-2 activity has been associated strongly with inflammatory diseases, cancer, and poor wound healing. The proposal is designed to develop compounds that can specifically inhibit MMP-2. Since cleavage by the MMP-2 requires binding of the substrate molecules, we propose to investigate the mechanism by which MMP-2 binds its main collagen substrate. In collaboration with other scientists, molecular biology and protein structural analyses are combined to first screen random peptide libraries for interaction with the collagen-binding domain (CBD) of MMP-2 and map the MMP-2 binding sites on collagen. Subsequent nuclear magnetic resonance structural studies will identify the collagen binding site on the MMP-2 using complexes containing the CBD from MMP-2 and collagen-like peptides. The specific interactions will be verified by introducing specific mutations into the identified binding site. Subsequently the bioactivity of the synthetic peptides will be enhanced by structure-based modifications. Subsequent experiments will determine whether the synthetic bioactive synthetic peptides and binding site regions may inhibit MMP-2 and alter the behavior of cells in cultures. The studies will integrate novel proteomics approaches to fully understand the mechanisms by which such peptides and binding site regions influence cell behavior. The proposed studies should define the specific binding sites of MMP-2 and collagen and explore a new strategy to inhibit MMP-2 in periodontal disease and oral cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Scientist Development Award - Research (K02)
Project #
1K02DE016312-01
Application #
6851316
Study Section
NIDCR Special Grants Review Committee (DSR)
Program Officer
Hardwick, Kevin S
Project Start
2004-09-28
Project End
2009-08-31
Budget Start
2004-09-28
Budget End
2005-08-31
Support Year
1
Fiscal Year
2004
Total Cost
$107,227
Indirect Cost
Name
University of Texas Health Science Center San Antonio
Department
Dentistry
Type
Schools of Dentistry
DUNS #
800772162
City
San Antonio
State
TX
Country
United States
Zip Code
78229
Mikhailova, Margarita; Xu, Xiaoping; Robichaud, Trista K et al. (2012) Identification of collagen binding domain residues that govern catalytic activities of matrix metalloproteinase-2 (MMP-2). Matrix Biol 31:380-8
Xu, Xiaoping; Mikhailova, Margarita; Chen, Zhihua et al. (2011) Peptide from the C-terminal domain of tissue inhibitor of matrix metalloproteinases-2 (TIMP-2) inhibits membrane activation of matrix metalloproteinase-2 (MMP-2). Matrix Biol 30:404-12
Steffensen, Bjorn; Chen, Zhihua; Pal, Sanjay et al. (2011) Fragmentation of fibronectin by inherent autolytic and matrix metalloproteinase activities. Matrix Biol 30:34-42
Pal, S; Chen, Z; Xu, X et al. (2010) Co-purified gelatinases alter the stability and biological activities of human plasma fibronectin preparations. J Periodontal Res 45:292-5
Jacobsen, Jasper N; Steffensen, Bjørn; Häkkinen, Lari et al. (2010) Skin wound healing in diabetic ?6 integrin-deficient mice. APMIS 118:753-64
Xu, Xiaoping; Mikhailova, Margarita; Ilangovan, Udayar et al. (2009) Nuclear magnetic resonance mapping and functional confirmation of the collagen binding sites of matrix metalloproteinase-2. Biochemistry 48:5822-31
Johnson, Dwight L; Carnes, David; Steffensen, Bjorn et al. (2009) Cellular effects of enamel matrix derivative are associated with different molecular weight fractions following separation by size-exclusion chromatography. J Periodontol 80:648-56
Stanley, Corey M; Wang, Yao; Pal, Sanjay et al. (2008) Fibronectin fragmentation is a feature of periodontal disease sites and diabetic foot and leg wounds and modifies cell behavior. J Periodontol 79:861-75
Murillo, Jesse; Wang, Yao; Xu, Xiaoping et al. (2008) Advanced glycation of type I collagen and fibronectin modifies periodontal cell behavior. J Periodontol 79:2190-9
Cortez, Dolores M; Feldman, Marc D; Mummidi, Srinivas et al. (2007) IL-17 stimulates MMP-1 expression in primary human cardiac fibroblasts via p38 MAPK- and ERK1/2-dependent C/EBP-beta , NF-kappaB, and AP-1 activation. Am J Physiol Heart Circ Physiol 293:H3356-65

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