Abstract

This proposal examines the role of specific membrane proteolipids (Prs) in the calcification of the oral microorganism, Corynebacterium (Formerly Bacterionema) matruchotii. The experiments are based on the hypothesis that the presence of specific membrane proteins promote formation of phospholipid:Ca:Pi complexes (CPLX), subsequent hydroxyapatite (HA) deposition, and serve a structural role in the formation of ion channels. Proteolipids which support HA formation have an Mr of 10,000 on SDS-PAGE and when recombined with other C. matruchotii proteolipids, ion transport is observed.
The Specific Aims are: 1.) To characterize the Mr 1O,OOO Pr(s) required for RA formation in vitro. 2. To compare the immunoreactive determinants of C. matruchotii Prs with those of other bacteria and the ability to calcify. 3.) To isolate Pr gene(s) using antibody probes. 4.) To examine the regulation of membrane mediated Pr-dependent HA formation in g. matruchotii. While our primary interest in this proposal is in microbial calcification, the technique developed and the principles established are directly relevant to membrane-mediated proteolipid-dependent calcification in vertebrate during endochondral ossification, dentinogenesis, or calcific disease. Biochemistry, immunology, and molecular biology will be used as tools to characterize the chemical and functional properties of these proteins during HA formation. Calcification in vitro will be used as a screening assay to identify and purify the 10K Pr that promotes HA deposition. Antibodies to this protein will be used to identify characteristics the promote HA formation via blocking experiments and by comparative analysis of immunoreactive determinants in CPLX. The antibody will also be used to clone the gene for this protein and to affinity purify the Pr. By varying culture physiology, the process of CPLX formation can be dissected, providing important new information applicable to septic stone a calculus formation as well as the basic process of calcification.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
2R01DE005932-08A3
Application #
3219655
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1981-01-01
Project End
1995-07-31
Budget Start
1990-08-01
Budget End
1991-07-31
Support Year
8
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
University of Texas Health Science Center San Antonio
Department
Type
Schools of Medicine
DUNS #
800772162
City
San Antonio
State
TX
Country
United States
Zip Code
78229

  Publications

Foster, Brian L; Popowics, Tracy E; Fong, Hanson K et al. (2007) Advances in defining regulators of cementum development and periodontal regeneration. Curr Top Dev Biol 78:47-126
Cohen, David J; Malave, David; Ghidoni, John J et al. (2004) Role of oral bacterial flora in calcific aortic stenosis: an animal model. Ann Thorac Surg 77:537-43
van Dijk, S; Dean, D D; Liu, Y et al. (1998) Purification, amino acid sequence, and cDNA sequence of a novel calcium-precipitating proteolipid involved in calcification of corynebacterium matruchotii. Calcif Tissue Int 62:350-8
Dean, D D; Schwartz, Z; Muniz, O E et al. (1992) Matrix vesicles are enriched in metalloproteinases that degrade proteoglycans. Calcif Tissue Int 50:342-9
Boskey, A L; Rimnac, C M; Bansal, M et al. (1992) Effect of short-term hypomagnesemia on the chemical and mechanical properties of rat bone. J Orthop Res 10:774-83
Dean, D D; Schwartz, Z V; Muniz, O E et al. (1992) Matrix vesicles contain metalloproteinases that degrade proteoglycans. Bone Miner 17:172-6
Boyan, B D; Schwartz, Z; Swain, L D (1992) In vitro studies on the regulation of endochondral ossification by vitamin D. Crit Rev Oral Biol Med 3:15-30
Boyan, B D; Schwartz, Z; Bonewald, L F et al. (1989) Localization of 1,25-(OH)2D3-responsive alkaline phosphatase in osteoblast-like cells (ROS 17/2.8, MG 63, and MC 3T3) and growth cartilage cells in culture. J Biol Chem 264:11879-86
Swain, L D; Renthal, R D; Boyan, B D (1989) Resolution of ion translocating proteolipid subclasses active in bacterial calcification. J Dent Res 68:1094-7
Boyan, B D; Schwartz, Z; Swain, L D et al. (1989) Role of lipids in calcification of cartilage. Anat Rec 224:211-9

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