Interaction of small heat shock proteins with light component of neurofilaments (NFL)
- 184 Downloads
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.
KeywordsSmall heat shock proteins Neurofilaments Cytoskeleton Intermediate filaments Neurodegenerative diseases
Small heat shock proteins
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).
- Ackerley S, James PA, Kalli A, French S, Davies KE, Talbot K (2006) A mutation in the small heat-shock protein HSPB1 leading to distal hereditary motor neuronopathy disrupts neurofilament assembly and the axonal transport of specific cellular cargoes. Hum Mol Genet 15(2):347–354. doi: 10.1093/hmg/ddi452 CrossRefPubMedGoogle Scholar
- Chalova AS, Sudnitsyna MV, Strelkov SV, Gusev NB (2014) Characterization of human small heat shock protein HspB1 that carries C-terminal domain mutations associated with hereditary motor neuron diseases. Biochim Biophys Acta 1844(12):2116–2126. doi: 10.1016/j.bbapap.2014.09.005 CrossRefPubMedGoogle Scholar
- Elliott JL, Der Perng M, Prescott AR, Jansen KA, Koenderink GH, Quinlan RA (2013) The specificity of the interaction between alphaB-crystallin and desmin filaments and its impact on filament aggregation and cell viability. Philos Trans R Soc Lond Ser B Biol Sci 368(1617):20120375. doi: 10.1098/rstb.2012.0375 CrossRefGoogle Scholar
- Evgrafov OV, Mersiyanova I, Irobi J, Van Den Bosch L, Dierick I, Leung CL, Schagina O, Verpoorten N, Van Impe K, Fedotov V, Dadali E, Auer-Grumbach M, Windpassinger C, Wagner K, Mitrovic Z, Hilton-Jones D, Talbot K, Martin JJ, Vasserman N, Tverskaya S, Polyakov A, Liem RK, Gettemans J, Robberecht W, De Jonghe P, Timmerman V (2004) Mutant small heat-shock protein 27 causes axonal Charcot-Marie-Tooth disease and distal hereditary motor neuropathy. Nat Genet 36(6):602–606. doi: 10.1038/ng1354 CrossRefPubMedGoogle Scholar
- Kayser J, Haslbeck M, Dempfle L, Krause M, Grashoff C, Buchner J, Herrmann H, Bausch AR (2013) The small heat shock protein Hsp27 affects assembly dynamics and structure of keratin intermediate filament networks. Biophys J 105(8):1778–1785. doi: 10.1016/j.bpj.2013.09.007 CrossRefPubMedPubMedCentralGoogle Scholar
- Muranova LK, Weeks SD, Strelkov SV, Gusev NB (2015) Characterization of mutants of human small heat shock protein HspB1 carrying replacements in the N-terminal domain and associated with hereditary motor neuron diseases. PLoS One 10(5):e0126248. doi: 10.1371/journal.pone.0126248 CrossRefPubMedPubMedCentralGoogle Scholar
- Nefedova VV, Datskevich PN, Sudnitsyna MV, Strelkov SV, Gusev NB (2013a) Physico-chemical properties of R140G and K141Q mutants of human small heat shock protein HspB1 associated with hereditary peripheral neuropathies. Biochimie 95(8):1582–1592. doi: 10.1016/j.biochi.2013.04.014 CrossRefPubMedGoogle Scholar