Amelogenin: Solution- and solid-state studies of a natural biomineralization process
EMSL Project ID
48235
Abstract
The protein amelogenin is the central protein controlling 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. We request EMSL resources to accomplish three major tasks. (1) In situ-AFM and related studies will allow us to physically view the oligomeric state of amelogenin bound to hydroxyapatite (HAP) surfaces. (2) Solution-state structural studies on full-length amelogen and LRAP will enable us to characterize the structure of these proteins under the variety of conditions found during enamel growth. (3) Solid-state NMR studies will enable the quantitative structural characterization of LRAP bound to hydroxyapatite. The AFM studies will involve two naturally occurring mutant amelogenin proteins observed in amelogenesis imperfecta. The solution-state NMR structural studies will focus on a Val, Leu, and Ile (?1) methyl-protonated 15N-, 13C-, 2H-labeled amelogenin sample. Such a labeling scheme in combination with transverse relaxation-optimized spectroscopy (TROSY) experiments, will allow us to track the structural behavior of amelogenin as it self-assembles into large molecular weight complexes. The solid-state NMR studies will focus on LRAP because 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
2013-12-20
End Date
2014-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Solid-State NMR Identification of Intermolecular Interactions in Amelogenin Bound to Hydroxyapatite
Jayasinha Arachchige R.M., S.D. Burton, J. Lu, B. Ginovska, L.K. Harding, M.E. Taylor, and J. Tao, et al. 2018. "Solid-State NMR Identification of Intermolecular Interactions in Amelogenin Bound to Hydroxyapatite." Biophysical Journal 115, no. 9:1666-1672. PNNL-SA-131727. doi:10.1016/j.bpj.2018.08.027
Secondary structure and dynamics study of the intrinsically disordered silica-mineralizing peptide P5S3during silicic acid condensation and silica decondensation
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
Sequence-Defined Energetic Shifts Control the Disassembly Kinetics and Microstructure of Amelogenin Adsorbed onto Hydroxyapatite (100)
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
The leucine-rich amelogenin protein (LRAP) is primarily monomeric and unstructured in physiological solution
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