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  • Chemistry  (2)
  • Analytical Chemistry and Spectroscopy  (1)
  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 8 (1990), S. 23-29 
    ISSN: 0887-3585
    Keywords: urea induced unfolding ; increased stability ; site-directed mutagenesis ; calbindin D9k ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: To investigate the contribution to protein stability of electrostatic interactions between charged surface residues, we have studied the effect of substituting three negatively charged solvent exposed residues with their side-chain amide analogs in bovine calbindin D9k - a small (Mr 8,500) globular protein of the calmodulin superfamily. The free energy of urea-induced unfolding for the wildtype and seven mutant proteins has been measured. The mutant proteins have increased stability towards unfolding relative to the wildtype. The experimental results correlate reasonably well with theoretically calculated relative free energies of unfolding and show that electrostatic interactions between charges on the surface of a protein can have significant effects on protein stability.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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  • 2
    ISSN: 0749-1581
    Keywords: 1H NMR ; 15N NMR ; Calbindin D9k ; Cadmium ; Calcium ; EF-hand calcium-binding proteins ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The amide 15N and 1H chemical shifts of (Cd2+)2-calbindin D9k are presented. A detailed comparison is made with those of (Ca2+)2-calbindin D9k, showing that the (Cd2+)2 and (Ca2+)2 states of the protein are very similar, with differences predominantly located in ion-binding loop I. The present studies were carried out on the P43G mutant calbindin D9k, and corroborate previous conclusions based on a comparative analysis of the 1H chemical shifts of the (Cd2+)2 and (Ca2+)2 states of wild-type calbindin D9k. A value for the rate constant for the dissociation of cadmium from ion-binding loop I in calbindin D9k of koff ≍ 0.6 × 103 ± 0.2 × 103 was estimated by analyzing line broadening in protein samples containing equal amounts of (Cd2+)1- and (Cd2+)2-calbindin D9k.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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