This proposal will explore novel structural principles by which proteins interact with their targets and will investigates the prevalence of "intrinsically disordered" proteins (IDPs) in the field of tooth biomineralization. The long term objective of the proposed research is to advance understanding of the role of extracellular matrix protein fragmentation in enamel and dentin, with a focus on the analysis of folding and the degree of disorder in the secondary structures of key proteins and polypeptides. We propose that disordered domains in the extracellular matrix proteins play substantial roles in biomineralization. We hypothesize that the programmed proteolytic activities in enamel and dentin regulate protein-mineral, protein-protein and protein- cell interactions through regulation of the number of unstructured regions and the degree of disorder in the protein sequence. Such interactions will affect chemical and cellular events such as cell signaling, macromolecular self-assembly, crystal nucleation and growth, and protein removal.
Three specific aims are proposed:
Aim I. To apply computational and biophysical strategies to analyze the secondary structures and evaluate the degree and nature of "disorder" in key proteins of the extracellular matrix of enamel and dentin as well as their selected proteolytic fragments.
Aim II : To use circular dichroism (CD) and fluorescence titrations to determine the conformational changes and strength of interactions between key fragments of amelogenin and ameloblastin, as well as DPP with their macromolecular targets. To use NMR spectroscopy to monitor key amino acid residues that undergoes conformational transitions in amelogenin sequence as the result of binding to partners/targets.
Aim III : To use spectroscopical techniques (CD and ATR-FTIR) to monitor secondary structural changes and conformational transitions of selected amelogenin, enamelin, and ameloblastin proteolytic fragments, as well as DPP following their interactions with calcium and with apatite crystal surfaces. In Summary: Enamel and dentin biomineralization is the result of orchestration among a series of protein-protein, protein-mineral and protein cell interactions. Our goal is to systematically dissect the sequence and secondary structures of functional domains in major enamel extracellular matrix proteins as well as the C-terminal portion of dentin sialophosphoprotein (DPP) with regard to IDPs, and to provide information on secondary structural alteration as the result of protein-protein, protein-mineral and protein-cell surface interactions. Understanding the role of "intrinsic disorder" in proteins of extracellular matrix of enamel and dentin will prepare the ground for the fabrication and development of biomimetic materials when synthetic peptides can be used to control the processes of crystal nucleation and growth. Identification of unfolded functional domains in cell signaling will have a great impact in the field of tissue regeneration. The outcomes of our study will therefore have the potential to improve treatments for repair and regeneration of oral, dental and craniofacial tissues.

Public Health Relevance

This proposal will explore novel structural principles by which proteins interact with their targets and will investigates the prevalence of "intrinsically disordered" proteins (IDPs) in the field of tooth biomineralization. The outcomes will prepare the ground for the fabrication and development of biomimetic materials when synthetic peptides can be used to control the processes of crystal nucleation and growth. Identification of unfolded functional domains in cell signaling will have a great impact in the field of tissue regeneration. The outcomes of our study will therefore have the potential to improve treatments for repair and regeneration of oral, dental and craniofacial tissues.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE020099-04
Application #
8306583
Study Section
Special Emphasis Panel (ZRG1-MOSS-K (09))
Program Officer
Lumelsky, Nadya L
Project Start
2009-09-25
Project End
2014-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
4
Fiscal Year
2012
Total Cost
$396,941
Indirect Cost
$151,916
Name
University of Southern California
Department
Dentistry
Type
Schools of Dentistry
DUNS #
072933393
City
Los Angeles
State
CA
Country
United States
Zip Code
90089
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