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Proline hydroxylase 2 (PHD2) promotes brown adipose thermogenesis by enhancing the hydroxylation of UCP1

OBJECTIVE: Brown adipose tissue (BAT) plays a crucial role in regulating non-shivering thermogenesis under cold exposure. Proline hydroxylases (PHDs) were found to be involved in adipocyte differentiation and lipid deposition. However, the effects of PHDs on regulatory mechanisms of BAT thermogenesi...

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Autores principales: Li, Fan, Zhang, Fenglin, Yi, Xin, Quan, Lu Lu, Yang, Xiaohua, Yin, Cong, Ma, Zewei, Wu, Ruifan, Zhao, Weijie, Ling, Mingfa, Lang, Limin, Hussein, Abdelaziz, Feng, Shengchun, Fu, Yiming, Wang, Junfeng, Liang, Shuyi, Zhu, Canjun, Wang, Lina, Zhu, Xiaotong, Gao, Ping, Xi, Qianyun, Zhang, Yongliang, Zhang, Lin, Shu, Gang, Jiang, Qingyan, Wang, Songbo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10293773/
https://www.ncbi.nlm.nih.gov/pubmed/37279828
http://dx.doi.org/10.1016/j.molmet.2023.101747
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author Li, Fan
Zhang, Fenglin
Yi, Xin
Quan, Lu Lu
Yang, Xiaohua
Yin, Cong
Ma, Zewei
Wu, Ruifan
Zhao, Weijie
Ling, Mingfa
Lang, Limin
Hussein, Abdelaziz
Feng, Shengchun
Fu, Yiming
Wang, Junfeng
Liang, Shuyi
Zhu, Canjun
Wang, Lina
Zhu, Xiaotong
Gao, Ping
Xi, Qianyun
Zhang, Yongliang
Zhang, Lin
Shu, Gang
Jiang, Qingyan
Wang, Songbo
author_facet Li, Fan
Zhang, Fenglin
Yi, Xin
Quan, Lu Lu
Yang, Xiaohua
Yin, Cong
Ma, Zewei
Wu, Ruifan
Zhao, Weijie
Ling, Mingfa
Lang, Limin
Hussein, Abdelaziz
Feng, Shengchun
Fu, Yiming
Wang, Junfeng
Liang, Shuyi
Zhu, Canjun
Wang, Lina
Zhu, Xiaotong
Gao, Ping
Xi, Qianyun
Zhang, Yongliang
Zhang, Lin
Shu, Gang
Jiang, Qingyan
Wang, Songbo
author_sort Li, Fan
collection PubMed
description OBJECTIVE: Brown adipose tissue (BAT) plays a crucial role in regulating non-shivering thermogenesis under cold exposure. Proline hydroxylases (PHDs) were found to be involved in adipocyte differentiation and lipid deposition. However, the effects of PHDs on regulatory mechanisms of BAT thermogenesis are not fully understood. METHODS: We detected the expression of PHDs in different adipose tissues by using immunoblotting and real-time PCR. Further, immunoblotting, real-time PCR, and immunostaining were performed to determine the correlation between proline hydroxylase 2 (PHD2) and UCP1 expression. Inhibitor of PHDs and PHD2-sgRNA viruses were used to construct the PHD2-deficiency model in vivo and in vitro to investigate the impacts of PHD2 on BAT thermogenesis. Afterward, the interaction between UCP1 and PHD2 and the hydroxylation modification level of UCP1 were verified by Co-IP assays and immunoblotting. Finally, the effect of specific proline hydroxylation on the expression/activity of UCP1 was further confirmed by site-directed mutation of UCP1 and mass spectrometry analysis. RESULTS: PHD2, but not PHD1 and PHD3, was highly enriched in BAT, colocalized, and positively correlated with UCP1. Inhibition or knockdown of PHD2 significantly suppressed BAT thermogenesis under cold exposure and aggravated obesity of mice fed HFD. Mechanistically, mitochondrial PHD2 bound to UCP1 and regulated the hydroxylation level of UCP1, which was enhanced by thermogenic activation and attenuated by PHD2 knockdown. Furthermore, PHD2-dependent hydroxylation of UCP1 promoted the expression and stability of UCP1 protein. Mutation of the specific prolines (Pro-33, 133, and 232) in UCP1 significantly mitigated the PHD2-elevated UCP1 hydroxylation level and reversed the PHD2-increased UCP1 stability. CONCLUSIONS: This study suggested an important role for PHD2 in BAT thermogenesis regulation by enhancing the hydroxylation of UCP1.
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spelling pubmed-102937732023-06-28 Proline hydroxylase 2 (PHD2) promotes brown adipose thermogenesis by enhancing the hydroxylation of UCP1 Li, Fan Zhang, Fenglin Yi, Xin Quan, Lu Lu Yang, Xiaohua Yin, Cong Ma, Zewei Wu, Ruifan Zhao, Weijie Ling, Mingfa Lang, Limin Hussein, Abdelaziz Feng, Shengchun Fu, Yiming Wang, Junfeng Liang, Shuyi Zhu, Canjun Wang, Lina Zhu, Xiaotong Gao, Ping Xi, Qianyun Zhang, Yongliang Zhang, Lin Shu, Gang Jiang, Qingyan Wang, Songbo Mol Metab Original Article OBJECTIVE: Brown adipose tissue (BAT) plays a crucial role in regulating non-shivering thermogenesis under cold exposure. Proline hydroxylases (PHDs) were found to be involved in adipocyte differentiation and lipid deposition. However, the effects of PHDs on regulatory mechanisms of BAT thermogenesis are not fully understood. METHODS: We detected the expression of PHDs in different adipose tissues by using immunoblotting and real-time PCR. Further, immunoblotting, real-time PCR, and immunostaining were performed to determine the correlation between proline hydroxylase 2 (PHD2) and UCP1 expression. Inhibitor of PHDs and PHD2-sgRNA viruses were used to construct the PHD2-deficiency model in vivo and in vitro to investigate the impacts of PHD2 on BAT thermogenesis. Afterward, the interaction between UCP1 and PHD2 and the hydroxylation modification level of UCP1 were verified by Co-IP assays and immunoblotting. Finally, the effect of specific proline hydroxylation on the expression/activity of UCP1 was further confirmed by site-directed mutation of UCP1 and mass spectrometry analysis. RESULTS: PHD2, but not PHD1 and PHD3, was highly enriched in BAT, colocalized, and positively correlated with UCP1. Inhibition or knockdown of PHD2 significantly suppressed BAT thermogenesis under cold exposure and aggravated obesity of mice fed HFD. Mechanistically, mitochondrial PHD2 bound to UCP1 and regulated the hydroxylation level of UCP1, which was enhanced by thermogenic activation and attenuated by PHD2 knockdown. Furthermore, PHD2-dependent hydroxylation of UCP1 promoted the expression and stability of UCP1 protein. Mutation of the specific prolines (Pro-33, 133, and 232) in UCP1 significantly mitigated the PHD2-elevated UCP1 hydroxylation level and reversed the PHD2-increased UCP1 stability. CONCLUSIONS: This study suggested an important role for PHD2 in BAT thermogenesis regulation by enhancing the hydroxylation of UCP1. Elsevier 2023-06-04 /pmc/articles/PMC10293773/ /pubmed/37279828 http://dx.doi.org/10.1016/j.molmet.2023.101747 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Li, Fan
Zhang, Fenglin
Yi, Xin
Quan, Lu Lu
Yang, Xiaohua
Yin, Cong
Ma, Zewei
Wu, Ruifan
Zhao, Weijie
Ling, Mingfa
Lang, Limin
Hussein, Abdelaziz
Feng, Shengchun
Fu, Yiming
Wang, Junfeng
Liang, Shuyi
Zhu, Canjun
Wang, Lina
Zhu, Xiaotong
Gao, Ping
Xi, Qianyun
Zhang, Yongliang
Zhang, Lin
Shu, Gang
Jiang, Qingyan
Wang, Songbo
Proline hydroxylase 2 (PHD2) promotes brown adipose thermogenesis by enhancing the hydroxylation of UCP1
title Proline hydroxylase 2 (PHD2) promotes brown adipose thermogenesis by enhancing the hydroxylation of UCP1
title_full Proline hydroxylase 2 (PHD2) promotes brown adipose thermogenesis by enhancing the hydroxylation of UCP1
title_fullStr Proline hydroxylase 2 (PHD2) promotes brown adipose thermogenesis by enhancing the hydroxylation of UCP1
title_full_unstemmed Proline hydroxylase 2 (PHD2) promotes brown adipose thermogenesis by enhancing the hydroxylation of UCP1
title_short Proline hydroxylase 2 (PHD2) promotes brown adipose thermogenesis by enhancing the hydroxylation of UCP1
title_sort proline hydroxylase 2 (phd2) promotes brown adipose thermogenesis by enhancing the hydroxylation of ucp1
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10293773/
https://www.ncbi.nlm.nih.gov/pubmed/37279828
http://dx.doi.org/10.1016/j.molmet.2023.101747
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