Unfolding and refolding of a quinone oxidoreductase: α-crystallin, a molecular chaperone, assists its reactivation

Goenka, Shradha ; Raman, Bakthisaran ; Ramakrishna, Tangirala ; Rao, Ch. Mohan (2001) Unfolding and refolding of a quinone oxidoreductase: α-crystallin, a molecular chaperone, assists its reactivation Biochemical journal, 359 . pp. 547-556. ISSN 0264-6021

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Official URL: http://www.biochemjusa.org/bj/359/bj3590547.htm

Abstract

α-Crystallin, a member of the small heat-shock protein family and present in vertebrate eye lens, is known to prevent the aggregation of other proteins under conditions of stress. However, its role in the reactivation of enzymes from their non-native inactive states has not been clearly demonstrated. We have studied the effect of α-crystallin on the refolding of ζ-crystallin, a quinone oxidoreductase, from its different urea-denatured states. Co-refolding ζ-crystallin from its denatured state in 2.5 M urea with either calf eye lens α-crystallin or recombinant human αB-crystallin could significantly enhance its reactivation yield. αB-crystallin was found to be more efficient than αA-crystallin in chaperoning the refolding of ζ-crystallin. In order to understand the nature of the denatured state(s) of ζ-crystallin that can interact with α-crystallin, we have investigated the unfolding pathway of ζ-crystallin. We find that it unfolds through three distinct intermediates: an altered tetramer, a partially unfolded dimer, which is competent to fold back to its active state, and a partially unfolded monomer. The partially unfolded monomer is inactive, exhibits highly exposed hydrophobic surfaces and has significant secondary structural elements with little or no tertiary structure. This intermediate does not refold into the active state without assistance. α-Crystallin provides the required assistance and improves the reactivation yield several-fold.

Item Type:Article
Source:Copyright of this article belongs to Portland Press.
Keywords:Chaperone-like Activity; ζ-Crystallin; Molten Globule, Reactivation; Unfolding
ID Code:35515
Deposited On:16 Apr 2011 14:10
Last Modified:17 May 2016 18:27

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