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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Published in:FEBS Open Bio
Main Authors: Frøbert, Anne Mette, Brohus, Malene, Toews, Julia N C, Round, Phillip, Fröbert, Ole, Hammond, Geoffrey L, Overgaard, Michael T
Format: Article in Journal/Newspaper
Language:English
Published: 2022
Subjects:
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
id ftalborgunivpubl:oai:pure.atira.dk:publications/0e2f5fbc-f98f-4dce-b12c-2b0bcda1d7e4
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spelling 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
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