Characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin
Sex hormone‐binding globulin (SHBG) regulates the bioavailability of sex steroid hormones in the blood. Levels of SHBG increase markedly in brown bears ( Ursus arctos ) during hibernation, suggesting that a key regulatory role of this protein is to quench sex steroid bioavailability in hibernation p...
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crwiley:10.1002/2211-5463.13341 2024-06-02T08:15:37+00:00 Characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin Frøbert, Anne Mette Brohus, Malene Toews, Julia N. C. Round, Phillip Fröbert, Ole Hammond, Geoffrey L. Overgaard, Michael T. Lundbeckfonden 2021 http://dx.doi.org/10.1002/2211-5463.13341 https://onlinelibrary.wiley.com/doi/pdf/10.1002/2211-5463.13341 https://onlinelibrary.wiley.com/doi/full-xml/10.1002/2211-5463.13341 https://febs.onlinelibrary.wiley.com/doi/pdf/10.1002/2211-5463.13341 en eng Wiley http://creativecommons.org/licenses/by/4.0/ FEBS Open Bio volume 12, issue 2, page 362-378 ISSN 2211-5463 2211-5463 journal-article 2021 crwiley https://doi.org/10.1002/2211-5463.13341 2024-05-03T11:26:03Z Sex hormone‐binding globulin (SHBG) regulates the bioavailability of sex steroid hormones in the blood. Levels of SHBG increase markedly in brown bears ( Ursus arctos ) during hibernation, suggesting that a key regulatory role of this protein is to quench sex steroid bioavailability in hibernation physiology. To enable characterization of ursine SHBG and a cross species comparison, we established an insect cell‐based expression system for recombinant full‐length ursine and human SHBG. Compared with human SHBG, we observed markedly lower secretion levels of ursine SHBG, resulting in a 10‐fold difference in purified protein yield. Both human and ursine recombinant SHBG appeared as dimeric proteins in solution, with a single unfolding temperature of ~ 58 °C. The thermal stability of ursine and human SHBG increased 5.4 and 9.5 °C, respectively, in the presence of dihydrotestosterone (DHT), suggesting a difference in affinity. The dissociation constants for [ 3 H]DHT were determined to 0.21 ± 0.04 n m for human and 1.32 ± 0.10 n m for ursine SHBG, confirming a lower affinity of ursine SHBG. A similarly reduced affinity, determined from competitive steroid binding, was observed for most steroids. Overall, we found that ursine SHBG had similar characteristics to human SHBG, specifically, being a homodimeric glycoprotein capable of binding steroids with high affinity. Therefore, ursine SHBG likely has similar biological functions to those known for human SHBG. The determined properties of ursine SHBG will contribute to elucidating its potential regulatory role in hibernation physiology. Article in Journal/Newspaper Ursus arctos Wiley Online Library FEBS Open Bio 12 2 362 378 |
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Wiley Online Library |
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crwiley |
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English |
description |
Sex hormone‐binding globulin (SHBG) regulates the bioavailability of sex steroid hormones in the blood. Levels of SHBG increase markedly in brown bears ( Ursus arctos ) during hibernation, suggesting that a key regulatory role of this protein is to quench sex steroid bioavailability in hibernation physiology. To enable characterization of ursine SHBG and a cross species comparison, we established an insect cell‐based expression system for recombinant full‐length ursine and human SHBG. Compared with human SHBG, we observed markedly lower secretion levels of ursine SHBG, resulting in a 10‐fold difference in purified protein yield. Both human and ursine recombinant SHBG appeared as dimeric proteins in solution, with a single unfolding temperature of ~ 58 °C. The thermal stability of ursine and human SHBG increased 5.4 and 9.5 °C, respectively, in the presence of dihydrotestosterone (DHT), suggesting a difference in affinity. The dissociation constants for [ 3 H]DHT were determined to 0.21 ± 0.04 n m for human and 1.32 ± 0.10 n m for ursine SHBG, confirming a lower affinity of ursine SHBG. A similarly reduced affinity, determined from competitive steroid binding, was observed for most steroids. Overall, we found that ursine SHBG had similar characteristics to human SHBG, specifically, being a homodimeric glycoprotein capable of binding steroids with high affinity. Therefore, ursine SHBG likely has similar biological functions to those known for human SHBG. The determined properties of ursine SHBG will contribute to elucidating its potential regulatory role in hibernation physiology. |
author2 |
Lundbeckfonden |
format |
Article in Journal/Newspaper |
author |
Frøbert, Anne Mette Brohus, Malene Toews, Julia N. C. Round, Phillip Fröbert, Ole Hammond, Geoffrey L. Overgaard, Michael T. |
spellingShingle |
Frøbert, Anne Mette Brohus, Malene Toews, Julia N. C. Round, Phillip Fröbert, Ole Hammond, Geoffrey L. Overgaard, Michael T. Characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin |
author_facet |
Frøbert, Anne Mette Brohus, Malene Toews, Julia N. C. Round, Phillip Fröbert, Ole Hammond, Geoffrey L. Overgaard, Michael T. |
author_sort |
Frøbert, Anne Mette |
title |
Characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin |
title_short |
Characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin |
title_full |
Characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin |
title_fullStr |
Characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin |
title_full_unstemmed |
Characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin |
title_sort |
characterization and comparison of recombinant full‐length ursine and human sex hormone‐binding globulin |
publisher |
Wiley |
publishDate |
2021 |
url |
http://dx.doi.org/10.1002/2211-5463.13341 https://onlinelibrary.wiley.com/doi/pdf/10.1002/2211-5463.13341 https://onlinelibrary.wiley.com/doi/full-xml/10.1002/2211-5463.13341 https://febs.onlinelibrary.wiley.com/doi/pdf/10.1002/2211-5463.13341 |
genre |
Ursus arctos |
genre_facet |
Ursus arctos |
op_source |
FEBS Open Bio volume 12, issue 2, page 362-378 ISSN 2211-5463 2211-5463 |
op_rights |
http://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.1002/2211-5463.13341 |
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FEBS Open Bio |
container_volume |
12 |
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2 |
container_start_page |
362 |
op_container_end_page |
378 |
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1800739849970384896 |