Defining the ligand-dependent proximatome of the sigma 1 receptor
Sigma 1 Receptor (S1R) is a therapeutic target for a wide spectrum of pathological conditions ranging from neurodegenerative diseases to cancer and COVID-19. S1R is ubiquitously expressed throughout the visceral organs, nervous, immune and cardiovascular systems. It is proposed to function as a liga...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-06-01
|
| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2023.1045759/full |
| _version_ | 1827931733008318464 |
|---|---|
| author | Jing Zhao Jing Zhao Rajalakshmi Veeranan-Karmegam Frederick C. Baker Barbara A. Mysona Barbara A. Mysona Barbara A. Mysona Pritha Bagchi Yutao Liu Yutao Liu Sylvia B. Smith Sylvia B. Smith Sylvia B. Smith Graydon B. Gonsalvez Kathryn E. Bollinger Kathryn E. Bollinger Kathryn E. Bollinger |
| author_facet | Jing Zhao Jing Zhao Rajalakshmi Veeranan-Karmegam Frederick C. Baker Barbara A. Mysona Barbara A. Mysona Barbara A. Mysona Pritha Bagchi Yutao Liu Yutao Liu Sylvia B. Smith Sylvia B. Smith Sylvia B. Smith Graydon B. Gonsalvez Kathryn E. Bollinger Kathryn E. Bollinger Kathryn E. Bollinger |
| author_sort | Jing Zhao |
| collection | DOAJ |
| description | Sigma 1 Receptor (S1R) is a therapeutic target for a wide spectrum of pathological conditions ranging from neurodegenerative diseases to cancer and COVID-19. S1R is ubiquitously expressed throughout the visceral organs, nervous, immune and cardiovascular systems. It is proposed to function as a ligand-dependent molecular chaperone that modulates multiple intracellular signaling pathways. The purpose of this study was to define the S1R proximatome under native conditions and upon binding to well-characterized ligands. This was accomplished by fusing the biotin ligase, Apex2, to the C terminus of S1R. Cells stably expressing S1R-Apex or a GFP-Apex control were used to map proximal proteins. Biotinylated proteins were labeled under native conditions and in a ligand dependent manner, then purified and identified using quantitative mass spectrometry. Under native conditions, S1R biotinylates over 200 novel proteins, many of which localize within the endomembrane system (endoplasmic reticulum, Golgi, secretory vesicles) and function within the secretory pathway. Under conditions of cellular exposure to either S1R agonist or antagonist, results show enrichment of proteins integral to secretion, extracellular matrix formation, and cholesterol biosynthesis. Notably, Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) displays increased binding to S1R under conditions of treatment with Haloperidol, a well-known S1R antagonist; whereas Low density lipoprotein receptor (LDLR) binds more efficiently to S1R upon treatment with (+)-Pentazocine ((+)-PTZ), a classical S1R agonist. Furthermore, we demonstrate that the ligand bound state of S1R correlates with specific changes to the cellular secretome. Our results are consistent with the postulated role of S1R as an intracellular chaperone and further suggest important and novel functionalities related to secretion and cholesterol metabolism. |
| first_indexed | 2024-03-13T06:57:54Z |
| format | Article |
| id | doaj.art-5fa23cfe8ef44a56808b7703df49b7a6 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-03-13T06:57:54Z |
| publishDate | 2023-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-5fa23cfe8ef44a56808b7703df49b7a62023-06-07T05:27:14ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2023-06-011110.3389/fcell.2023.10457591045759Defining the ligand-dependent proximatome of the sigma 1 receptorJing Zhao0Jing Zhao1Rajalakshmi Veeranan-Karmegam2Frederick C. Baker3Barbara A. Mysona4Barbara A. Mysona5Barbara A. Mysona6Pritha Bagchi7Yutao Liu8Yutao Liu9Sylvia B. Smith10Sylvia B. Smith11Sylvia B. Smith12Graydon B. Gonsalvez13Kathryn E. Bollinger14Kathryn E. Bollinger15Kathryn E. Bollinger16Department of Ophthalmology, Medical College of Georgia at Augusta University, Augusta, GA, United StatesCulver Vision Discovery Institute, Augusta, GA, United StatesDepartment of Cellular Biology and Anatomy, Medical College of Georgia at Augusta University, Augusta, GA, United StatesDepartment of Cellular Biology and Anatomy, Medical College of Georgia at Augusta University, Augusta, GA, United StatesDepartment of Ophthalmology, Medical College of Georgia at Augusta University, Augusta, GA, United StatesCulver Vision Discovery Institute, Augusta, GA, United StatesDepartment of Cellular Biology and Anatomy, Medical College of Georgia at Augusta University, Augusta, GA, United StatesEmory Integrated Proteomics Core, Emory University, Atlanta, GA, United StatesCulver Vision Discovery Institute, Augusta, GA, United StatesDepartment of Cellular Biology and Anatomy, Medical College of Georgia at Augusta University, Augusta, GA, United StatesDepartment of Ophthalmology, Medical College of Georgia at Augusta University, Augusta, GA, United StatesCulver Vision Discovery Institute, Augusta, GA, United StatesDepartment of Cellular Biology and Anatomy, Medical College of Georgia at Augusta University, Augusta, GA, United StatesDepartment of Cellular Biology and Anatomy, Medical College of Georgia at Augusta University, Augusta, GA, United StatesDepartment of Ophthalmology, Medical College of Georgia at Augusta University, Augusta, GA, United StatesCulver Vision Discovery Institute, Augusta, GA, United StatesDepartment of Cellular Biology and Anatomy, Medical College of Georgia at Augusta University, Augusta, GA, United StatesSigma 1 Receptor (S1R) is a therapeutic target for a wide spectrum of pathological conditions ranging from neurodegenerative diseases to cancer and COVID-19. S1R is ubiquitously expressed throughout the visceral organs, nervous, immune and cardiovascular systems. It is proposed to function as a ligand-dependent molecular chaperone that modulates multiple intracellular signaling pathways. The purpose of this study was to define the S1R proximatome under native conditions and upon binding to well-characterized ligands. This was accomplished by fusing the biotin ligase, Apex2, to the C terminus of S1R. Cells stably expressing S1R-Apex or a GFP-Apex control were used to map proximal proteins. Biotinylated proteins were labeled under native conditions and in a ligand dependent manner, then purified and identified using quantitative mass spectrometry. Under native conditions, S1R biotinylates over 200 novel proteins, many of which localize within the endomembrane system (endoplasmic reticulum, Golgi, secretory vesicles) and function within the secretory pathway. Under conditions of cellular exposure to either S1R agonist or antagonist, results show enrichment of proteins integral to secretion, extracellular matrix formation, and cholesterol biosynthesis. Notably, Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) displays increased binding to S1R under conditions of treatment with Haloperidol, a well-known S1R antagonist; whereas Low density lipoprotein receptor (LDLR) binds more efficiently to S1R upon treatment with (+)-Pentazocine ((+)-PTZ), a classical S1R agonist. Furthermore, we demonstrate that the ligand bound state of S1R correlates with specific changes to the cellular secretome. Our results are consistent with the postulated role of S1R as an intracellular chaperone and further suggest important and novel functionalities related to secretion and cholesterol metabolism.https://www.frontiersin.org/articles/10.3389/fcell.2023.1045759/fullmolecular chapteroneERGICcholesterolPCSK9LDLRsecretome |
| spellingShingle | Jing Zhao Jing Zhao Rajalakshmi Veeranan-Karmegam Frederick C. Baker Barbara A. Mysona Barbara A. Mysona Barbara A. Mysona Pritha Bagchi Yutao Liu Yutao Liu Sylvia B. Smith Sylvia B. Smith Sylvia B. Smith Graydon B. Gonsalvez Kathryn E. Bollinger Kathryn E. Bollinger Kathryn E. Bollinger Defining the ligand-dependent proximatome of the sigma 1 receptor Frontiers in Cell and Developmental Biology molecular chapterone ERGIC cholesterol PCSK9 LDLR secretome |
| title | Defining the ligand-dependent proximatome of the sigma 1 receptor |
| title_full | Defining the ligand-dependent proximatome of the sigma 1 receptor |
| title_fullStr | Defining the ligand-dependent proximatome of the sigma 1 receptor |
| title_full_unstemmed | Defining the ligand-dependent proximatome of the sigma 1 receptor |
| title_short | Defining the ligand-dependent proximatome of the sigma 1 receptor |
| title_sort | defining the ligand dependent proximatome of the sigma 1 receptor |
| topic | molecular chapterone ERGIC cholesterol PCSK9 LDLR secretome |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2023.1045759/full |
| work_keys_str_mv | AT jingzhao definingtheliganddependentproximatomeofthesigma1receptor AT jingzhao definingtheliganddependentproximatomeofthesigma1receptor AT rajalakshmiveeranankarmegam definingtheliganddependentproximatomeofthesigma1receptor AT frederickcbaker definingtheliganddependentproximatomeofthesigma1receptor AT barbaraamysona definingtheliganddependentproximatomeofthesigma1receptor AT barbaraamysona definingtheliganddependentproximatomeofthesigma1receptor AT barbaraamysona definingtheliganddependentproximatomeofthesigma1receptor AT prithabagchi definingtheliganddependentproximatomeofthesigma1receptor AT yutaoliu definingtheliganddependentproximatomeofthesigma1receptor AT yutaoliu definingtheliganddependentproximatomeofthesigma1receptor AT sylviabsmith definingtheliganddependentproximatomeofthesigma1receptor AT sylviabsmith definingtheliganddependentproximatomeofthesigma1receptor AT sylviabsmith definingtheliganddependentproximatomeofthesigma1receptor AT graydonbgonsalvez definingtheliganddependentproximatomeofthesigma1receptor AT kathrynebollinger definingtheliganddependentproximatomeofthesigma1receptor AT kathrynebollinger definingtheliganddependentproximatomeofthesigma1receptor AT kathrynebollinger definingtheliganddependentproximatomeofthesigma1receptor |