Structure-function correlations of proteins in enamel formation
EMSL Project ID
19851a
Abstract
Heterogeneous nucleation is important in the development of biominerals such as bone, teeth and mollusk shells, but often, initial nucleation events are not understood at a mechanistic level. It is generally agreed that proteins play a vital role in the nucleation event, however, the specific protein is often not identified and protein structures are often not available. Our proposal would allow us to address some of the key structural questions for the biomineralization protein, amelogenin, both as a monomer and in its nanospherical functional form, and correlate the structure with the function of nucleation. Challenges to determine the solubilized structure previously have been the lack of the ability to crystallize the protein, and the functional form being a nanosphere composed of 20-100 monomers. We recently have had some success collecting and analyzing data for the monomer, but are limited by resolution at low field which would be enhanced with EMSL's high field NMR capabilities. The structure of the protein will be investigated using dynamic light scattering (DLS) and high field solution state NMR. The nucleation event will be investigated using SEM, XRD, TEM and ICP to characterize the resulting calcium phosphate particles. These results will provide critical new insights into the structure and the function of the amelogenin protein, with a larger impact on the formation and stabilization of biominerals in general and the nanoscale synthesis of materials in the materials community.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2007-06-01
End Date
2009-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Buchko GW, BJ Tarasevich, JG Bekhazi, ML Snead, and WJ Shaw. 2008. "A Solution NMR Investigation into the Early Events of Amelogenin Nanosphere Self-Assembly Initiated with Sodium Chloride or Calcium Chloride." Biochemistry 47(50):13215-13222. doi:10.1021/bi8018288
Buchko GW, JG Bekhazi, JR Cort, NB Valentine, ML Snead, and WJ Shaw. 2008. "1H, 13C, and 15N resonance assignments of murine amelogenin, an enamel biomineralization protein. ." Biomolecular NMR Assignments 2(1):89-91.
Buchko, GW, Tarasevich, BJ, Roberts, J, Snead, ML, and Shaw, WJ (2010) "A solution NMR investigation into the murine amelogenin splice-variant LRAP (Leucine-Rich Amelogenin Protein)." Biochim. Biophys. Acta ? Proteins and Proteomics, 1804:1768-1774.
doi:10.1016/j.bbapap.2010.03.006
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.
Tarasevich BJ, AS Lea, and WJ Shaw. 2010. "The leucine rich amelogenin protein (LRAP) adsorbs as monomers or dimers onto surfaces." Journal of Structural Biology 169(3):266-276. doi:10.1016/j.jsb.2009.10.007
Tarasevich BJ, AS Lea, W Bernt, MH Engelhard, and WJ Shaw. 2009. "Adsorption of Amelogenin onto Self-Assembled and Fluoroapatite Surfaces." Journal of Physical Chemistry B 113(7):1833-1842.
doi: 10.1021/jp804548x
Tarasevich BJ, AS Lea, W Bernt, MH Engelhard, and WJ Shaw. 2009. "Changes in the quaternary structure of amelogenin when adsorbed onto surfaces." Biopolymers 91(2):103-107. doi:10.1002/bip.21095
Tarasevich BJ, CJ Howard, JL Larson, ML Snead, J Simmer, ML Paine, and WJ Shaw. 2007. "The Nucleation and growth of Calcium Phosphate by Amelogenin." Journal of Crystal Growth 304(2):407-415.