This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Mineralized tissues are composites of organic (collagen, other proteins) and inorganic (mineral) components. Biomineralization of these tissues is closely controlled by non-collagenous proteins (NCPs) within the tissue organic matrix to produce biological materials with unique mechanical and biological properties tailored for their functions. The size, shape, orientation and location of mineral crystals is regulated by these proteins in order to achieve these remarkable properties. Control of biomineralization is incompletely understood, including the NCPs involved in the regulation in different tissues and their specific roles, changes that occur in diseases involving mineralization, and the physicochemical mechanisms employed by NCPs to control mineralization through direct NCP-mineral interactions. The proposed research aims to elucidate the molecular events of biomineralization through two project components. The first component is the development of a gel electrophoresis/diffusion system to facilitate fast analysis of collagen-based tissues for these mineralization regulators. The proposed gel system will be capable of separating small amounts of mineralization-regulating species from complex tissue matrices and evaluating their effect on mineralization in vitro. Once optimized, this gel system will be used to evaluate differences in NCP complements at different stages of mineralization, as well as in healthy versus diseased mineralized tissue. The second component of the project aims to investigate structure and conformation changes in NCPs at the interface of protein and mineral. Two structural techniques, circular dichroism (CD) spectrophotometry and solid-state nuclear magnetic resonance (NMR) will be employed to investigate the structure of NCP-derived peptides adsorbed to mineral crystals. From these investigations a model of the protein-mineral interaction will emerge, shedding light on the physicochemical mechanisms by which these proteins influence mineral growth in vivo.
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