αA-Crystallin-derived mini-chaperone modulates stability and function of cataract causing αAG98R-crystallin.
A substitution mutation in human αA-crystallin (αAG98R) is associated with autosomal dominant cataract. The recombinant mutant αAG98R protein exhibits altered structure, substrate-dependent chaperone activity, impaired oligomer stability and aggregation on prolonged incubation at 37 °C. Our previous...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3435407?pdf=render |
| _version_ | 1828802832476667904 |
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| author | Murugesan Raju Puttur Santhoshkumar K Krishna Sharma |
| author_facet | Murugesan Raju Puttur Santhoshkumar K Krishna Sharma |
| author_sort | Murugesan Raju |
| collection | DOAJ |
| description | A substitution mutation in human αA-crystallin (αAG98R) is associated with autosomal dominant cataract. The recombinant mutant αAG98R protein exhibits altered structure, substrate-dependent chaperone activity, impaired oligomer stability and aggregation on prolonged incubation at 37 °C. Our previous studies have shown that αA-crystallin-derived mini-chaperone (DFVIFLDVKHFSPEDLTVK) functions like a molecular chaperone by suppressing the aggregation of denaturing proteins. The present study was undertaken to determine the effect of αA-crystallin-derived mini-chaperone on the stability and chaperone activity of αAG98R-crystallin.Recombinant αAG98R was incubated in presence and absence of mini-chaperone and analyzed by chromatographic and spectrometric methods. Transmission electron microscope was used to examine the effect of mini-chaperone on the aggregation propensity of mutant protein. Mini-chaperone containing photoactive benzoylphenylalanine was used to confirm the interaction of mini-chaperone with αAG98R. The rescuing of chaperone activity in mutantα-crystallin (αAG98R) by mini-chaperone was confirmed by chaperone assays. We found that the addition of the mini-chaperone during incubation of αAG98R protected the mutant crystallin from forming larger aggregates that precipitate with time. The mini-chaperone-stabilized αAG98R displayed chaperone activity comparable to that of wild-type αA-crystallin. The complexes formed between mini-αA-αAG98R complex and ADH were more stable than the complexes formed between αAG98R and ADH. Western-blotting and mass spectrometry confirmed the binding of mini-chaperone to mutant crystallin.These results demonstrate that mini-chaperone stabilizes the mutant αA-crystallin and modulates the chaperone activity of αAG98R. These findings aid in our understanding of how to design peptide chaperones that can be used to stabilize mutant αA-crystallins and preserve the chaperone function. |
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| id | doaj.art-6bf238bc46ec4602bc167495e5d5d10b |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T07:11:19Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-6bf238bc46ec4602bc167495e5d5d10b2022-12-22T00:33:37ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0179e4407710.1371/journal.pone.0044077αA-Crystallin-derived mini-chaperone modulates stability and function of cataract causing αAG98R-crystallin.Murugesan RajuPuttur SanthoshkumarK Krishna SharmaA substitution mutation in human αA-crystallin (αAG98R) is associated with autosomal dominant cataract. The recombinant mutant αAG98R protein exhibits altered structure, substrate-dependent chaperone activity, impaired oligomer stability and aggregation on prolonged incubation at 37 °C. Our previous studies have shown that αA-crystallin-derived mini-chaperone (DFVIFLDVKHFSPEDLTVK) functions like a molecular chaperone by suppressing the aggregation of denaturing proteins. The present study was undertaken to determine the effect of αA-crystallin-derived mini-chaperone on the stability and chaperone activity of αAG98R-crystallin.Recombinant αAG98R was incubated in presence and absence of mini-chaperone and analyzed by chromatographic and spectrometric methods. Transmission electron microscope was used to examine the effect of mini-chaperone on the aggregation propensity of mutant protein. Mini-chaperone containing photoactive benzoylphenylalanine was used to confirm the interaction of mini-chaperone with αAG98R. The rescuing of chaperone activity in mutantα-crystallin (αAG98R) by mini-chaperone was confirmed by chaperone assays. We found that the addition of the mini-chaperone during incubation of αAG98R protected the mutant crystallin from forming larger aggregates that precipitate with time. The mini-chaperone-stabilized αAG98R displayed chaperone activity comparable to that of wild-type αA-crystallin. The complexes formed between mini-αA-αAG98R complex and ADH were more stable than the complexes formed between αAG98R and ADH. Western-blotting and mass spectrometry confirmed the binding of mini-chaperone to mutant crystallin.These results demonstrate that mini-chaperone stabilizes the mutant αA-crystallin and modulates the chaperone activity of αAG98R. These findings aid in our understanding of how to design peptide chaperones that can be used to stabilize mutant αA-crystallins and preserve the chaperone function.http://europepmc.org/articles/PMC3435407?pdf=render |
| spellingShingle | Murugesan Raju Puttur Santhoshkumar K Krishna Sharma αA-Crystallin-derived mini-chaperone modulates stability and function of cataract causing αAG98R-crystallin. PLoS ONE |
| title | αA-Crystallin-derived mini-chaperone modulates stability and function of cataract causing αAG98R-crystallin. |
| title_full | αA-Crystallin-derived mini-chaperone modulates stability and function of cataract causing αAG98R-crystallin. |
| title_fullStr | αA-Crystallin-derived mini-chaperone modulates stability and function of cataract causing αAG98R-crystallin. |
| title_full_unstemmed | αA-Crystallin-derived mini-chaperone modulates stability and function of cataract causing αAG98R-crystallin. |
| title_short | αA-Crystallin-derived mini-chaperone modulates stability and function of cataract causing αAG98R-crystallin. |
| title_sort | αa crystallin derived mini chaperone modulates stability and function of cataract causing αag98r crystallin |
| url | http://europepmc.org/articles/PMC3435407?pdf=render |
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