roles of electrostatics and conformation in protein-crystal interactions静电学和构象的角色在测定晶体的相互作用.pdfVIP

Roles of Electrostatics and Conformation in Protein-Crystal Interactions 1 2 3 4 1 Paul V. Azzopardi , Jason O’Young , Gilles Lajoie , Mikko Karttunen , Harvey A. Goldberg , Graeme K. Hunter1* 1 School of Dentistry and Department of Biochemistry, University of Western Ontario, London, Ontario, Canada, 2 School of Dentistry, University of Western Ontario, London, Ontario, Canada, 3 Department of Biochemistry, University of Western Ontario, London, Ontario, Canada, 4 Department of Applied Mathematics, University of Western Ontario, London, Ontario, Canada Abstract In vitro studies have shown that the phosphoprotein osteopontin (OPN) inhibits the nucleation and growth of hydroxyapatite (HA) and other biominerals. In vivo, OPN is believed to prevent the calcification of soft tissues. However, the nature of the interaction between OPN and HA is not understood. In the computational part of the present study, we used molecular dynamics simulations to predict the adsorption of 19 peptides, each 16 amino acids long and collectively covering the entire sequence of OPN, to the {100} face of HA. This analysis showed that there is an inverse relationship between predicted strength of adsorption and peptide isoelectric point (P,0.0001). Analysis of the OPN sequence by PONDR (Predictor of Naturally Disordered Regions) indicated that OPN sequences predicted to adsorb well to HA are highly disordered. In the experimental part of the study, we synthesized phosphorylated and non-phosphorylated peptides corresponding to OPN sequences 65–80 (pSHDHMDDDDDDDDDGD) and 220–235 (pSHEpSTEQSDAIDpSAEK). In agreement with the PONDR analysis, these were shown by circular