Shifting the competition between the intramolecular reshuffling reaction and the direct oxidation reaction during the oxidative folding of kinetically trapped disulfide-insecure intermediates

Mahesh Narayan, E. Welker, Celestine Wanjalla, Guoqiang Xu, Harold A. Scheraga

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The oxidative folding pathway(s) of single-domain proteins can be characterized by the existence, stability, and structural nature of the intermediates that populate the regeneration pathway. Structured intermediates can be disulfide-secure in that they are able to protect their existing (native) disulfide bonds from SH/SS reshuffling and reduction reactions, and thereby form the native protein directly, i.e., by oxidation of their exposed (or locally exposable) thiols. Alternatively, they can be disulfide-insecure, usually requiring global unfolding to expose their free thiols. However, such an unfolding event also exposes the existing native disulfide bonds. Thus, the subsequent oxidation reaction to form the native protein in a disulfide-insecure intermediate competes with the intramolecular attack by the thiols of the macromolecule on its own native disulfide bonds, resulting in a large population of intermediates that are reshuffled instead of being oxidized. Under stabilizing conditions, disulfide-insecure species become long-lived kinetically trapped intermediates that slowly and only indirectly convert to the native protein through reshuffling reactions. In this study, trans-[Pt(en)2Cl2]2+, a strong oxidizing agent which has not traditionally been used in oxidative folding, was applied to shift the competition between reshuffling and oxidation reactions in des [58-110] and des [26-84], two long-lived disulfide-insecure intermediates of RNase A, and oxidize them directly under stable conditions to form the native protein. Such a successful direct conversion of kinetically trapped intermediates to the native molecule by trans-[Pt(en)2Cl 2]2+ may be helpful in facilitating the oxidative folding processes of multi-disulfide-containing proteins in general.

Original languageEnglish
Pages (from-to)10783-10789
Number of pages7
JournalBiochemistry
Volume42
Issue number36
DOIs
Publication statusPublished - Sep 16 2003

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Disulfides
Oxidation
Sulfhydryl Compounds
Proteins
Pancreatic Ribonuclease
Macromolecules
Oxidants
Regeneration
Molecules
Population

ASJC Scopus subject areas

  • Biochemistry

Cite this

Shifting the competition between the intramolecular reshuffling reaction and the direct oxidation reaction during the oxidative folding of kinetically trapped disulfide-insecure intermediates. / Narayan, Mahesh; Welker, E.; Wanjalla, Celestine; Xu, Guoqiang; Scheraga, Harold A.

In: Biochemistry, Vol. 42, No. 36, 16.09.2003, p. 10783-10789.

Research output: Contribution to journalArticle

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