Enamel biomineralization: Solution- and solid-state NMR studies of a natural protein-mineral interface
EMSL Project ID
47735
Abstract
The protein amelogenin is the central protein that controls the formation of dental enamel, the hardest tissue in the human body. The protein orchestrates the nucleation, growth, and organization of enamel formation (amelogenesis) through the formation of large quaternary structures called nanospheres, consisting of 20-100 monomers. The structure and function of full-length amelogenin and LRAP, the latter a splice variant of amelogenin that contains only the charged N- and C- termini, are the focus of our four-year, NIH funded investigations. Solution-state structural studies are being conducted on full-length amelogen, two naturally occurring mutants, and LRAP, with an emphasis on characterizing structure under the variety of conditions found during enamel growth. Solid-state quantitative structural and orientation studies of the protein bound to hydroxyapatite are focused on LRAP, as its smaller size allows easy incorporation of selective isotopic labels which enable the determination of site specific, molecular level protein structure, protein-surface interactions and dynamics. Structural changes observed in solution or in the solid-state will then be correlated to differences in function.
Project Details
Start Date
2012-12-05
End Date
2013-11-13
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Buchko GW, G Lin, BJ Tarasevich, and WJ Shaw. 2013. "A solution NMR investigation into the impaired self-assembly properties of two murine amelogenins containing the point mutations T21?I or P41?T." Archives of Biochemistry and Biophysics 537(2):217-224. doi:10.1016/j.abb.2013.07.015
G.W. Buchko and W.J. Shaw (2015) Improved protocol to purify untagged amelogenin - Application to murine amelogenin containing the equivalent P70&8594T point mutation observed in human amelogenesis imperfecta. Protein Expr. Purif.. 105:14-22.
Lu J, Y Xu, GW Buchko, and WJ Shaw. 2013. "Mineral Association Changes the Secondary Structure and Dynamics of Murine Amelogenin." Journal of Dental Research 92(11):1000-1004. doi:10.1177/0022034513504929
Tao J, GW Buchko, WJ Shaw, J De Yoreo, and BJ Tarasevich. 2015. "Sequence-defined Energetic Shifts Control the Disassembly Kinetics and Microstructure of Amelogenin Adsorbed onto Hydroxyapatite (100)." Langmuir 31(38):10451-10460. doi:10.1021/acs.langmuir.5b02549
Tarasevich BJ, JS Philo, NK Maluf, S Krueger, GW Buchko, G Lin, and WJ Shaw. 2015. "The Leucine-Rich Amelogenin Protein (LRAP) is primarily monomeric and unstructured in physiological solution." Journal of Structural Biology 190(1):81-91. doi:10.1016/j.jsb.2014.10.007
Zerfab C, GW Buchko, WJ Shaw, S Hobe, and H Paulsen. 2017. "Secondary structure and dynamics study of the intrinsically disordered silica-mineralizing peptide P5S3 during silicic acid condensation and silica decondensation." Proteins. Structure, Function, and Bioinformatics 85(11):2111-2126. doi:10.1002/prot.25366