Mutations in Human αA-crystallin/sHSP Affect Subunit Exchange Interaction with αB-crystallin

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2012 Feb 21

PMID 22347476

Citations 8

Authors

Ilangovan Raju

Lalita Oonthonpan

Edathara C Abraham

Affiliations

Soon will be listed here.

Abstract

Background: Mutation in αA-crystallin contributes to the development of congenital cataract in humans. Heterooligomerization of αA-crystallin and αB-crystallin is essential for maintaining transparency in the eye lens. The effect of congenital cataract causing mutants of αA-crystallin on subunit exchange and interaction with αB-crystallin is unknown. In the present study, interaction of the mutants of αA-crystallin with αB-crystallin was studied both in vitro and in situ by the fluorescence resonance energy transfer (FRET) technique.

Methodology/principal Findings: In vitro FRET technique was used to demonstrate the rates of subunit exchange of αB-wt with the following αA-crystallin mutants: R12C, R21L, R21W, R49C, R54C, and R116C. The subunit exchange rates (k values) of R21W and R116C with αB-wt decreased drastically as compared to αA-wt interacting with αB-wt. Moderately decreased k values were seen with R12C, R49C and R54C while R21L showed nearly normal k value. The interaction of αA- mutants with αB-wt was also assessed by in situ FRET. YFP-tagged αA mutants were co-expressed with CFP-tagged αB-wt in HeLa cells and the spectral signals were captured with a confocal microscope before and after acceptor laser photobleaching. The interaction of R21W and R116C with αB-wt was decreased nearly 50% as compared to αA-wt while the rest of the mutants showed slightly decreased interaction. Thus, there is good agreement between the in vitro and in situ FRET data.

Conclusions/significance: Structural changes occurring in these mutants, as reported earlier, could be the underlying cause for the decreased interaction with αB may contribute to development of congenital cataract.

Citing Articles

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Congenital cataract - clinical and morphological aspects.

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A novel mutation in the CRYAA gene associated with congenital cataract and microphthalmia in a Chinese family.

Song Z, Si N, Xiao W BMC Med Genet. 2018; 19(1):190.

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