Complete sequence-specific, proton-resonance assignments have been determined for the calcium phosphate-stabilizing tryptic peptide β-casein-(1–25) containing the phosphorylated sequence motif Ser(P)17-Ser(P)-Ser(P)-Glu-Glu21. Spectra of the peptide have been recorded, in separate experiments, in the presence of excess ammonium ions, sodium ions and calcium ions, and of the dephosphorylated peptide in the presence of excess sodium ions. We observed significant changes to chemical shifts for backbone and side-chain resonances that were dependent upon the nature of the cation present. Medium-range nuclear Overhauser effect (nOe) enhancements, characteristic of small structured regions in the peptide, were observed and also found to be cation dependent. The secondary structure of the peptide was characterized by sequential and medium-range (i, i+2/3/4, which denotes an interaction between residue i and residue i+2, i+3 or i+4 in the peptide) nOe connectivities, and Hα chemical shifts. Four structured regions were identified in the calcium-bound peptide: residues Arg1 to Glu4 were involved in a loop-type structure, and residues Val8 to Glu11, Ser(P)17 to Glu20 and Glu21 to Thr24 were implicated in β-turn conformations. Comparison of the patterns of medium-range nOe connectivities in β-casein-(1–25) with those in αS1-casein-(59–79) suggest that the two peptides have distinctly different conformations in the presence of calcium ions, despite having a high degree of sequential and functional similarity.
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Research Article|
May 08 2001
Cation-dependent structural features of β-casein-(1–25)
Keith J. CROSS;
Keith J. CROSS
∗School of Dental Science, The University of Melbourne, 711 Elizabeth Street, Melbourne, Victoria 3000, Australia
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N. Laila HUQ;
N. Laila HUQ
∗School of Dental Science, The University of Melbourne, 711 Elizabeth Street, Melbourne, Victoria 3000, Australia
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Wendy BICKNELL;
Wendy BICKNELL
†Victorian College of Pharmacy, Royal Parade, Parkville, Victoria 3052, Australia
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Eric C. REYNOLDS
Eric C. REYNOLDS
1
∗School of Dental Science, The University of Melbourne, 711 Elizabeth Street, Melbourne, Victoria 3000, Australia
1To whom correspondence should be addressed (e-mail e.reynolds@unimelb.edu.au).
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Publisher: Portland Press Ltd
Received:
October 11 2000
Revision Received:
February 02 2001
Accepted:
March 09 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2001
2001
Biochem J (2001) 356 (1): 277–286.
Article history
Received:
October 11 2000
Revision Received:
February 02 2001
Accepted:
March 09 2001
Citation
Keith J. CROSS, N. Laila HUQ, Wendy BICKNELL, Eric C. REYNOLDS; Cation-dependent structural features of β-casein-(1–25). Biochem J 15 May 2001; 356 (1): 277–286. doi: https://doi.org/10.1042/bj3560277
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