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Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B
Room-temperature X-ray crystallography provides unique insights into protein conformational heterogeneity, but obtaining sufficiently large protein crystals is a common hurdle. Serial synchrotron crystallography (SSX) helps to address this hurdle by allowing the use of many medium- to small-sized cr...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9813971/ https://www.ncbi.nlm.nih.gov/pubmed/36598353 http://dx.doi.org/10.1107/S2053230X22011645 |
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author | Sharma, Shivani Ebrahim, Ali Keedy, Daniel A. |
author_facet | Sharma, Shivani Ebrahim, Ali Keedy, Daniel A. |
author_sort | Sharma, Shivani |
collection | PubMed |
description | Room-temperature X-ray crystallography provides unique insights into protein conformational heterogeneity, but obtaining sufficiently large protein crystals is a common hurdle. Serial synchrotron crystallography (SSX) helps to address this hurdle by allowing the use of many medium- to small-sized crystals. Here, a recently introduced serial sample-support chip system has been used to obtain the first SSX structure of a human phosphatase, specifically protein tyrosine phosphatase 1B (PTP1B) in the unliganded (apo) state. In previous apo room-temperature structures, the active site and allosteric sites adopted alternate conformations, including open and closed conformations of the active-site WPD loop and of a distal allosteric site. By contrast, in our SSX structure the active site is best fitted with a single conformation, but the distal allosteric site is best fitted with alternate conformations. This observation argues for additional nuance in interpreting the nature of allosteric coupling in this protein. Overall, our results illustrate the promise of serial methods for room-temperature crystallography, as well as future avant-garde crystallography experiments, for PTP1B and other proteins. |
format | Online Article Text |
id | pubmed-9813971 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-98139712023-01-09 Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B Sharma, Shivani Ebrahim, Ali Keedy, Daniel A. Acta Crystallogr F Struct Biol Commun Research Communications Room-temperature X-ray crystallography provides unique insights into protein conformational heterogeneity, but obtaining sufficiently large protein crystals is a common hurdle. Serial synchrotron crystallography (SSX) helps to address this hurdle by allowing the use of many medium- to small-sized crystals. Here, a recently introduced serial sample-support chip system has been used to obtain the first SSX structure of a human phosphatase, specifically protein tyrosine phosphatase 1B (PTP1B) in the unliganded (apo) state. In previous apo room-temperature structures, the active site and allosteric sites adopted alternate conformations, including open and closed conformations of the active-site WPD loop and of a distal allosteric site. By contrast, in our SSX structure the active site is best fitted with a single conformation, but the distal allosteric site is best fitted with alternate conformations. This observation argues for additional nuance in interpreting the nature of allosteric coupling in this protein. Overall, our results illustrate the promise of serial methods for room-temperature crystallography, as well as future avant-garde crystallography experiments, for PTP1B and other proteins. International Union of Crystallography 2023-01-01 /pmc/articles/PMC9813971/ /pubmed/36598353 http://dx.doi.org/10.1107/S2053230X22011645 Text en © Shivani Sharma et al. 2023 https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Communications Sharma, Shivani Ebrahim, Ali Keedy, Daniel A. Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B |
title | Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B |
title_full | Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B |
title_fullStr | Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B |
title_full_unstemmed | Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B |
title_short | Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B |
title_sort | room-temperature serial synchrotron crystallography of the human phosphatase ptp1b |
topic | Research Communications |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9813971/ https://www.ncbi.nlm.nih.gov/pubmed/36598353 http://dx.doi.org/10.1107/S2053230X22011645 |
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