The Heterooligomerization of Human Small Heat Shock Proteins Is Controlled by Conserved Motif Located in the N-Terminal Domain
| Autor: | Nikolai B. Gusev, Vladislav M Shatov, Sergei V. Strelkov |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Chemistry Multidisciplinary Amino Acid Motifs Mutant Ion chromatography OLIGOMERIZATION Pentapeptide repeat MEMBER lcsh:Chemistry Molar ratio lcsh:QH301-705.5 Spectroscopy Chemistry small heat shock proteins HspB8 General Medicine HspB6 Computer Science Applications HspB1 HspB5 Physical Sciences Chromatography Gel COMPLEXES Life Sciences & Biomedicine heterooligomerization FORM Biochemistry & Molecular Biology animal structures Stereochemistry ALPHA-B-CRYSTALLIN Article Catalysis Inorganic Chemistry 03 medical and health sciences HSP22 HSPB8 Protein Domains Humans Physical and Theoretical Chemistry Molecular Biology Small Heat-Shock Proteins Science & Technology 030102 biochemistry & molecular biology SEQUENCES Conserved motif Organic Chemistry Heat-Shock Proteins Small 030104 developmental biology lcsh:Biology (General) lcsh:QD1-999 Mutation Protein Multimerization |
| Zdroj: | International Journal of Molecular Sciences Volume 21 Issue 12 International Journal of Molecular Sciences, Vol 21, Iss 4248, p 4248 (2020) |
| Popis: | Ubiquitously expressed human small heat shock proteins (sHsps) HspB1, HspB5, HspB6 and HspB8 contain a conserved motif (S/G)RLFD in their N-terminal domain. For each of them, we prepared mutants with a replacement of the conserved R by A (R/A mutants) and a complete deletion of the pentapeptide (&Delta mutants) and analyzed their heterooligomerization with other wild-type (WT) human sHsps. We found that WT HspB1 and HspB5 formed heterooligomers with HspB6 only upon heating. In contrast, both HspB1 mutants interacted with WT HspB6 even at low temperature. HspB1/HspB6 heterooligomers revealed a broad size distribution with equimolar ratio suggestive of heterodimers as building blocks, while HspB5/HspB6 heterooligomers had an approximate 2:1 ratio. In contrast, R/A or &Delta mutants of HspB6, when mixed with either HspB1 or HspB5, resulted in heterooligomers with a highly variable molar ratio and a decreased HspB6 incorporation. No heterooligomerization of HspB8 or its mutants with either HspB1 or HspB5 could be detected. Finally, R/A or &Delta mutations had no effect on heterooligomerization of HspB1 and HspB5 as analyzed by ion exchange chromatography. We conclude that the conserved N-terminal motif plays an important role in heterooligomer formation, as especially pronounced in HspB6 lacking the C-terminal IXI motif. |
| Databáze: | OpenAIRE |
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