Solid State NMR Investigation of the Structure of Statherin Protein on the Surface of Hydroxyapatite
EMSL Project ID
7601
Abstract
The structure of proteins involved in engineering and maintenance of hard tissue is of great importance to the understanding of aging processes, injury recovery mechanisms and pathological calcification pathways. The salivary protein, statherin, which assumes an important role in biomineralization processes has been studied using solid state NMR experiments on the surface of the mineral hydroxyapatite (HAP). The first 15 residues in the protein N-terminus were shown to posses a rigid helical structure from structural and dynamics measurements [1-5]. For the study of the rest of the protein, the following labeling scheme was devised, based on protein folding models, generated by David Baker?s powerful prediction engine [6]. DpSpSEEKFLRRIGRFGYGY[2-13C]GPYQ[4?-19F]PVPEQPLYPQ[1-13C]P[1-13C]YQPQ[15N]YQQYTF
The models were statistically analyzed to narrow down the choices of labeling positions in the protein so as to probe potential long-range and short-range interactions in the intermediate region and the C-terminus region, respectively. Site-specific isotopic labeling at five specific locations of the full-length protein was carried out using solid phase peptide synthesis.
Structural investigations of the protein on the surface of HAP will be carried out using MAS NMR measurements. In particular, DRAWS and REDOR experiments will be used to examine a predicted helical structure in the C-terminus of the protein and to detect distances of P23, a suspected hinge residue, to both the C- and the N-terminus.
Project Details
Project type
Capability Research
Start Date
2004-09-04
End Date
2005-09-06
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members