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Cell Stress and Chaperones

, Volume 22, Issue 4, pp 467–479 | Cite as

Interaction of small heat shock proteins with light component of neurofilaments (NFL)

  • Victoria V. Nefedova
  • Maria V. Sudnitsyna
  • Nikolai B. Gusev
SMALL HEAT SHOCK PROTEINS

Abstract

The interaction of human small heat shock protein HspB1, its point mutants associated with distal hereditary motor neuropathy, and three other small heat shock proteins (HspB5, HspB6, HspB8) with the light component of neurofilaments (NFL) was analyzed by differential centrifugation, analytical ultracentrifugation, and fluorescent spectroscopy. The wild-type HspB1 decreased the quantity of NFL in pellets obtained after low- and high-speed centrifugation and increased the quantity of NFL remaining in the supernatant after high-speed centrifugation. Part of HspB1 was detected in the pellet of NFL after high-speed centrifugation, and at saturation, 1 mol of HspB1 monomer was bound per 2 mol of NFL. Point mutants of HspB1 associated with distal hereditary motor neuropathy (G84R, L99M, R140G, K141Q, and P182S) were almost as effective as the wild-type HspB1 in modulation of NFL assembly. At low ionic strength, HspB1 weakly interacted with NFL tetramers, and this interaction was increased upon salt-induced polymerization of NFL. HspB1 and HspB5 (αB-crystallin) decreased the rate of NFL polymerization measured by fluorescent spectroscopy. HspB6 (Hsp20) and HspB8 (Hsp22) were less effective than HspB1 (or HspB5) in modulation of NFL assembly. The data presented indicate that the small heat shock proteins affect NFL transition from tetramers to filaments, hydrodynamic properties of filaments, and their bundling and therefore probably modulate the formation of intermediate filament networks in neurons.

Keywords

Small heat shock proteins Neurofilaments Cytoskeleton Intermediate filaments Neurodegenerative diseases 

Abbreviations

DTT

Dithiothreitol

IFs

Intermediate filaments

IPTG

Isopropyl β-D-1-thiogalactopyranoside

ME

Mercaptoethanol

SEC

Size-exclusion chromatography

sHsp

Small heat shock proteins

TCA

Trichloroacetic acid

Notes

Acknowledgements

The authors are grateful to Dr. Oleg Klychnikov (Laboratory for Biocrystallography, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Belgium) for the valuable discussion and linguistic advice. Investigation of the effect of HspB1 mutants on NFL assembly was supported by grant 16-04-00016 of the Russian Foundation for Basic Science, and investigation of different small heat shock proteins was supported by grant 14-35-00026 of the Russian Science Foundation (RSF).

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Copyright information

© Cell Stress Society International 2016

Authors and Affiliations

  • Victoria V. Nefedova
    • 1
  • Maria V. Sudnitsyna
    • 1
  • Nikolai B. Gusev
    • 1
  1. 1.Department of Biochemistry, School of BiologyMoscow State UniversityMoscowRussian Federation

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