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On the reactivity of carboxyl groups of ribonuclease-A in anhydrous methanol

Acharya, AS and Vithayathil, PJ (1975) On the reactivity of carboxyl groups of ribonuclease-A in anhydrous methanol. In: International Journal of Peptide & Protein Research, 7 (3). pp. 207-219.

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Official URL: http://onlinelibrary.wiley.com/doi/10.1111/j.1399-...


The esterification of Ribonuclease-A in methanol/0.1 M hydrochloric acid has been studied by measuring the decrease in the number of titratable groups of the protein and estimating the amount of methanol incorporated. Esterification of nearly five of the 11 free carboxyl groups of the protein resulted in almost complete inactivation of the enzyme. The initial products of esterification have been chromatographed on Amberlite columns, and five partially active methyl ester derivatives of Ribonuclease-A have been isolated. The dimethyl ester, the initial product of esterification with reduced catalytic activity, has the carboxyl groups of Glu-49 and Asp-53 modified. Even in the non-aqueous solvent, as in the native structure of the protein in aqueous solution, these carboxyl groups are the fast reacting ones. Subsquently, the esterification reaction appears to proceed preferentially at the C-terminal region of the molecule. Comparison of the reactivities of carboxyl groups of Ribonuclease-A in acidic methanol to that known in aqueous solutions (with carbodiimides) suggests that the structure of Ribonuclease-A in the non-aqueous solvent resembles, at least in part, the structure in aqueous environment.

Item Type: Journal Article
Publication: International Journal of Peptide & Protein Research
Publisher: John Wiley and Sons
Additional Information: Copyright of this article belongs to John Wiley & Sons.
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 13 Jan 2010 09:54
Last Modified: 23 Feb 2012 06:50
URI: http://eprints.iisc.ac.in/id/eprint/24069

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