Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp)
In this study, a reliable and simple method of untagged recombinant human HspB7 preparation was developed. Recombinant HspB7 is presented in two oligomeric forms with an apparent molecular weight of 36 kDa (probably dimers) and oligomers with an apparent molecular weight of more than 600 kDa. By usi...
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| Format: | Article |
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/15/7777 |
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| author | Lydia K. Muranova Vladislav M. Shatov Andrey V. Slushchev Nikolai B. Gusev |
| author_facet | Lydia K. Muranova Vladislav M. Shatov Andrey V. Slushchev Nikolai B. Gusev |
| author_sort | Lydia K. Muranova |
| collection | DOAJ |
| description | In this study, a reliable and simple method of untagged recombinant human HspB7 preparation was developed. Recombinant HspB7 is presented in two oligomeric forms with an apparent molecular weight of 36 kDa (probably dimers) and oligomers with an apparent molecular weight of more than 600 kDa. By using hydrophobic and size-exclusion chromatography, we succeeded in preparation of HspB7 dimers. Mild oxidation promoted the formation of large oligomers, whereas the modification of Cys 126 by iodoacetamide prevented it. The deletion of the first 13 residues or deletion of the polySer motif (residues 17–29) also prevented the formation of large oligomers of HspB7. Cys-mutants of HspB6 and HspB8 containing a single-Cys residue in the central part of the β7 strand in a position homologous to that of Cys137 in HspB1 can be crosslinked to the wild-type HspB7 through a disulfide bond. Immobilized on monoclonal antibodies, the wild-type HspB6 interacted with the wild-type HspB7. We suppose that formation of heterodimers of HspB7 with HspB6 and HspB8 may be important for the functional activity of these small heat shock proteins. |
| first_indexed | 2024-03-10T09:15:49Z |
| format | Article |
| id | doaj.art-5674b5f85d144b6b8d04ead16ba53462 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T09:15:49Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
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| spelling | doaj.art-5674b5f85d144b6b8d04ead16ba534622023-11-22T05:38:11ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012215777710.3390/ijms22157777Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp)Lydia K. Muranova0Vladislav M. Shatov1Andrey V. Slushchev2Nikolai B. Gusev3Department of Biochemistry, School of Biology, Moscow State University, 19234 Moscow, RussiaDepartment of Biochemistry, School of Biology, Moscow State University, 19234 Moscow, RussiaDepartment of Biochemistry, School of Biology, Moscow State University, 19234 Moscow, RussiaDepartment of Biochemistry, School of Biology, Moscow State University, 19234 Moscow, RussiaIn this study, a reliable and simple method of untagged recombinant human HspB7 preparation was developed. Recombinant HspB7 is presented in two oligomeric forms with an apparent molecular weight of 36 kDa (probably dimers) and oligomers with an apparent molecular weight of more than 600 kDa. By using hydrophobic and size-exclusion chromatography, we succeeded in preparation of HspB7 dimers. Mild oxidation promoted the formation of large oligomers, whereas the modification of Cys 126 by iodoacetamide prevented it. The deletion of the first 13 residues or deletion of the polySer motif (residues 17–29) also prevented the formation of large oligomers of HspB7. Cys-mutants of HspB6 and HspB8 containing a single-Cys residue in the central part of the β7 strand in a position homologous to that of Cys137 in HspB1 can be crosslinked to the wild-type HspB7 through a disulfide bond. Immobilized on monoclonal antibodies, the wild-type HspB6 interacted with the wild-type HspB7. We suppose that formation of heterodimers of HspB7 with HspB6 and HspB8 may be important for the functional activity of these small heat shock proteins.https://www.mdpi.com/1422-0067/22/15/7777small heat shock proteinsHspB7cvHspoligomeric structure |
| spellingShingle | Lydia K. Muranova Vladislav M. Shatov Andrey V. Slushchev Nikolai B. Gusev Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp) International Journal of Molecular Sciences small heat shock proteins HspB7 cvHsp oligomeric structure |
| title | Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp) |
| title_full | Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp) |
| title_fullStr | Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp) |
| title_full_unstemmed | Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp) |
| title_short | Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp) |
| title_sort | quaternary structure and hetero oligomerization of recombinant human small heat shock protein hspb7 cvhsp |
| topic | small heat shock proteins HspB7 cvHsp oligomeric structure |
| url | https://www.mdpi.com/1422-0067/22/15/7777 |
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