ISSN:
0006-3525
Schlagwort(e):
Chemistry
;
Polymer and Materials Science
Quelle:
Wiley InterScience Backfile Collection 1832-2000
Thema:
Chemie und Pharmazie
Notizen:
The divalent cation-induced protein folding properties of the template macromolecule, bovine dentine phosphophoryn (BDPP), have been examined by 1H/31P/13C/113Cd-nmr spectroscopy. Cd(II) was employed, exploiting the sensitivity of 113Cd-nmr to ligand-binding interactions and kinetics. Cation binding was studied over the stoichiometric range of 0-50 : 1 Cd(II) : protein (mole ratio), well below the range of Cd(II) concentration required to induce protein precipitation. The stepwise titration of divalent cation-depleted phosphophoryn at pH 7.2 in H2O/D2O with 113CdCl2 revealed that (PSer)n, (PSerAsp)n, and (Asp)n polyelectrolyte cation-binding domains undergo two major transitions in their secondary and tertiary structures: the first transition, occurring between 1 : 10 and 1 : 1 Cd(II) : protein stoichiometry, and the second, between 10 : 1 and 50 : 1. By monitoring the amide NH intensities, 31P-nmr chemical shift, and 13C Asp-C, resonances, it was concluded that Cd(II) ions exhibit a binding-site preference for polyelectrolyte cation-binding domains, in the order \documentclass{article}\pagestyle{empty}\begin{document}$$ {\rm (PSer)}_n {\rm 〉 (PSerAsp)}_n {\rm 〉 (Asp)}_n $$\end{document} This preference correlates with the degree of negative charge density for each sequence motif. Accompanying the backbone conformational transitions at the polyelectrolyte regions were conformational transitions in the flanking hinge domains, indicating that the hinge domains participate in the folding of the phosphophoryn molecule as divalent cation binding occurs at the polyelectrolyte domains.We were unsuccessful in detecting phosphophoryn-bound Cd (11) species by 113Cd-nmr because of chemical exchange modulation. However, using a smaller 21-residue peptide mimetic of phosphophoryn, we have observed three stoichiometric-dependent 113Cd resonances that differ in terms of the oxoanion coordination number. Our observation of multiple Cd(II) species in the presence of the peptide supports our contention that Cd(II) has many chemically distinct coordination sites on phosphophoryn, each in multiple equilibria with H2O, Cl-, and side-chain oxoatoms. © 1994 John Wiley & Sons, Inc.
Zusätzliches Material:
9 Ill.
Materialart:
Digitale Medien
URL:
http://dx.doi.org/10.1002/bip.360341008
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