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 phy...
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2022
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Online Access: | https://vbn.aau.dk/da/publications/0e2f5fbc-f98f-4dce-b12c-2b0bcda1d7e4 https://doi.org/10.1002/2211-5463.13341 https://vbn.aau.dk/ws/files/469375880/FEBS_Open_Bio_2021_Fr_bert_Characterization_and_comparison_of_recombinant_full_length_ursine_and_human_sex.pdf http://www.scopus.com/inward/record.url?scp=85121041688&partnerID=8YFLogxK |
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ftalborgunivpubl:oai:pure.atira.dk:publications/0e2f5fbc-f98f-4dce-b12c-2b0bcda1d7e4 2024-09-15T18:40:15+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 2022-02 application/pdf https://vbn.aau.dk/da/publications/0e2f5fbc-f98f-4dce-b12c-2b0bcda1d7e4 https://doi.org/10.1002/2211-5463.13341 https://vbn.aau.dk/ws/files/469375880/FEBS_Open_Bio_2021_Fr_bert_Characterization_and_comparison_of_recombinant_full_length_ursine_and_human_sex.pdf http://www.scopus.com/inward/record.url?scp=85121041688&partnerID=8YFLogxK eng eng https://vbn.aau.dk/da/publications/0e2f5fbc-f98f-4dce-b12c-2b0bcda1d7e4 info:eu-repo/semantics/openAccess Frøbert , A M , Brohus , M , Toews , J N C , Round , P , Fröbert , O , Hammond , G L & Overgaard , M T 2022 , ' Characterization and comparison of recombinant full-length ursine and human sex hormone-binding globulin ' , FEBS open bio , vol. 12 , no. 2 , pp. 362-378 . https://doi.org/10.1002/2211-5463.13341 Animals Dihydrotestosterone/metabolism Humans Recombinant Proteins Sex Hormone-Binding Globulin/chemistry Steroids/metabolism Ursidae /dk/atira/pure/sustainabledevelopmentgoals/life_on_land name=SDG 15 - Life on Land /dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being name=SDG 3 - Good Health and Well-being article 2022 ftalborgunivpubl https://doi.org/10.1002/2211-5463.13341 2024-08-29T00:19:40Z 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 nm for human and 1.32 ± 0.10 nm 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 Aalborg University's Research Portal FEBS Open Bio 12 2 362 378 |
institution |
Open Polar |
collection |
Aalborg University's Research Portal |
op_collection_id |
ftalborgunivpubl |
language |
English |
topic |
Animals Dihydrotestosterone/metabolism Humans Recombinant Proteins Sex Hormone-Binding Globulin/chemistry Steroids/metabolism Ursidae /dk/atira/pure/sustainabledevelopmentgoals/life_on_land name=SDG 15 - Life on Land /dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being name=SDG 3 - Good Health and Well-being |
spellingShingle |
Animals Dihydrotestosterone/metabolism Humans Recombinant Proteins Sex Hormone-Binding Globulin/chemistry Steroids/metabolism Ursidae /dk/atira/pure/sustainabledevelopmentgoals/life_on_land name=SDG 15 - Life on Land /dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being name=SDG 3 - Good Health and Well-being 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 |
topic_facet |
Animals Dihydrotestosterone/metabolism Humans Recombinant Proteins Sex Hormone-Binding Globulin/chemistry Steroids/metabolism Ursidae /dk/atira/pure/sustainabledevelopmentgoals/life_on_land name=SDG 15 - Life on Land /dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being name=SDG 3 - Good Health and Well-being |
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 nm for human and 1.32 ± 0.10 nm 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. |
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 |
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 |
publishDate |
2022 |
url |
https://vbn.aau.dk/da/publications/0e2f5fbc-f98f-4dce-b12c-2b0bcda1d7e4 https://doi.org/10.1002/2211-5463.13341 https://vbn.aau.dk/ws/files/469375880/FEBS_Open_Bio_2021_Fr_bert_Characterization_and_comparison_of_recombinant_full_length_ursine_and_human_sex.pdf http://www.scopus.com/inward/record.url?scp=85121041688&partnerID=8YFLogxK |
genre |
Ursus arctos |
genre_facet |
Ursus arctos |
op_source |
Frøbert , A M , Brohus , M , Toews , J N C , Round , P , Fröbert , O , Hammond , G L & Overgaard , M T 2022 , ' Characterization and comparison of recombinant full-length ursine and human sex hormone-binding globulin ' , FEBS open bio , vol. 12 , no. 2 , pp. 362-378 . https://doi.org/10.1002/2211-5463.13341 |
op_relation |
https://vbn.aau.dk/da/publications/0e2f5fbc-f98f-4dce-b12c-2b0bcda1d7e4 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1002/2211-5463.13341 |
container_title |
FEBS Open Bio |
container_volume |
12 |
container_issue |
2 |
container_start_page |
362 |
op_container_end_page |
378 |
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1810484566579740672 |