The Heterooligomerization of Human Small Heat Shock Proteins Is Controlled by Conserved Motif Located in the N-Terminal Domain
Abstract
:1. Introduction
2. Results
2.1. Characterization of Analyzed Proteins
2.2. Effect of R/A and Δ Mutations on the Interaction of HspB1 and HspB6
2.3. Effect of R/A and Δ Mutations on the Interaction of HspB5 and HspB6
2.4. Lack of Interaction of HspB8 and Its Mutants with HspB1 and HspB5
2.5. Formation of Heterooligomeric Complexes between HspB1 and HspB5
3. Discussion
4. Material and Methods
4.1. Cloning of the Wild-Type Proteins and Their Mutants
4.2. Expression and Purification of Recombinant Proteins
4.3. Formation of Heterooligomeric Complexes
4.4. Analysis of Heterooligomeric Complexes
Author Contributions
Funding
Conflicts of Interest
Abbreviations
DTT | dithiothreitol |
ME | mercaptoethanol |
PMSF | phenylmethanesulphonyl fluoride |
SDS | sodium dodecyl sulfate |
SEC | size-exclusion chromatography |
TCA | trichloroacetic acid |
WT | wild type |
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WT HspB1 | R/A HspB1 | Δ HspB1 | WT HspB5 | R/A HspB5 | Δ HspB5 | ||
---|---|---|---|---|---|---|---|
WT HspB5 | Without heating | 0 | 0 | 0 | |||
After heating | 1.00 (constant ~1.0) | 1.00 (constant ~1.0) | 1.00 (constant ~1.0) | ||||
R/A HspB5 | Without heating | 0 | |||||
After heating | 1.00 (constant ~1.0) | ||||||
Δ HspB5 | Without heating | 0 | |||||
After heating | 1.00 (constant ~1.0) | ||||||
WT HspB6 | Without heating | 0.05 | 0.31 (variable 5–1) | 0.82 (variable 3–1) | 0.05 | 0.12 | 0 |
After heating | 0.99 (constant ~1.1) | 0.99 (constant ~1.1) | 1.00 (constant ~1.2) | 0.99 (constant ~2.0) | 0.99 (variable 3–2) | 0.98 (variable 5–2) | |
R/A HspB6 | Without heating | 0.04 | 0.12 | ||||
After heating | 0.78 (variable 10–1) | 0.99 (variable 4–1) | |||||
Δ HspB6 | Without heating | 0.05 | 0 | ||||
After heating | 0.32 (variable 10–1) | 0.98 (variable 4–1) | |||||
WT HspB8 | Without heating | 0 | 0 | 0 | 0 | 0 | 0 |
After heating | 0 | 0 | 0 | 0 | 0 | 0 | |
R/A HspB8 | Without heating | 0 | 0 | ||||
After heating | 0 | 0 | |||||
Δ HspB8 | Without heating | 0 | 0 | ||||
After heating | 0 | 0 |
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Shatov, V.M.; Strelkov, S.V.; Gusev, N.B. The Heterooligomerization of Human Small Heat Shock Proteins Is Controlled by Conserved Motif Located in the N-Terminal Domain. Int. J. Mol. Sci. 2020, 21, 4248. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21124248
Shatov VM, Strelkov SV, Gusev NB. The Heterooligomerization of Human Small Heat Shock Proteins Is Controlled by Conserved Motif Located in the N-Terminal Domain. International Journal of Molecular Sciences. 2020; 21(12):4248. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21124248
Chicago/Turabian StyleShatov, Vladislav M., Sergei V. Strelkov, and Nikolai B. Gusev. 2020. "The Heterooligomerization of Human Small Heat Shock Proteins Is Controlled by Conserved Motif Located in the N-Terminal Domain" International Journal of Molecular Sciences 21, no. 12: 4248. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21124248