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The pivotal role of the β7 strand in the intersubunit contacts of different human small heat shock proteins

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Cell Stress and Chaperones Aims and scope

Abstract

Human αB-crystallin and small heat shock proteins HspB6 and HspB8 were mutated so that all endogenous Cys residues were replaced by Ser and the single Cys residue was inserted in a position homologous to that of Cys137 of human HspB1, i.e. in a position presumably located in the central part of β7 strand of the α-crystallin domain. The secondary, tertiary, and quaternary structures of thus obtained Cys-mutants as well as their chaperone-like activity were similar to those of their wild-type counterparts. Mild oxidation of Cys-mutants leads to formation of disulfide bond crosslinking neighboring monomers thus indicating participation of the β7 strand in intersubunit interaction. Oxidation weakly affects the secondary and tertiary structure, does not affect the quaternary structure of αB-crystallin and HspB6, and shifts equilibrium between monomer and dimer of HspB8 towards dimer formation. It is concluded that the β7 strand participates in the intersubunit interaction of four human small heat shock proteins (αB-crystallin, HspB1, HspB6, HspB8) having different structure of β2 strand of α-crystallin domain and different length and composition of variable N- and C-terminal tails.

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Abbreviations

Cys-mutant of αB-crystallin:

Human αB-crystallin with point mutation E117C

Сys-mutant of HspB6:

Human HspB6 with double mutation C46S/E116C

Cys-mutant of HspB8:

Human HspB8 with fourfold mutation C10S/C99S/C195S/N138C

bis-ANS:

4,4′-Bis(1-anilinonaphthalene-8-sulfonate)

DTT:

Dithiothreitol

ME:

β-Mercaptoethanol

PMSF:

Phenylmethanesulfonyl fluoride

sHsp:

Small heat shock proteins

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Acknowledgment

This investigation was supported by the Russian Foundation for Basic Research.

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Authors and Affiliations

  1. Department of Biochemistry, School of Biology, Moscow State University, Moscow, 119991, Russian Federation

    Evgeny V. Mymrikov, Olesya V. Bukach, Alim S. Seit-Nebi & Nikolai B. Gusev

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  1. Evgeny V. Mymrikov
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  2. Olesya V. Bukach
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  3. Alim S. Seit-Nebi
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  4. Nikolai B. Gusev
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Correspondence to Nikolai B. Gusev.

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Mymrikov, E.V., Bukach, O.V., Seit-Nebi, A.S. et al. The pivotal role of the β7 strand in the intersubunit contacts of different human small heat shock proteins. Cell Stress and Chaperones 15, 365–377 (2010). https://doi.org/10.1007/s12192-009-0151-8

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  • DOI: https://doi.org/10.1007/s12192-009-0151-8

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