Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology]
Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers
The American Society for Biochemistry and Molecular Biology (ASBMB)
Published 2018
The American Society for Biochemistry and Molecular Biology (ASBMB)
Published 2018
Publication Date: |
2018-02-24
|
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Publisher: |
The American Society for Biochemistry and Molecular Biology (ASBMB)
|
Print ISSN: |
0021-9258
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Electronic ISSN: |
1083-351X
|
Topics: |
Biology
Chemistry and Pharmacology
|
Published by: |
_version_ | 1836398810416283648 |
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autor | Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers |
beschreibung | Neurosteroids are endogenous sterols that potentiate or inhibit pentameric ligand-gated ion channels (pLGICs) and can be effective anesthetics, analgesics, or anti-epileptic drugs. The complex effects of neurosteroids on pLGICs suggest the presence of multiple binding sites in these receptors. Here, using a series of novel neurosteroid-photolabeling reagents combined with top-down and middle-down mass spectrometry, we have determined the stoichiometry, sites, and orientation of binding for 3α,5α-pregnane neurosteroids in the Gloeobacter ligand-gated ion channel (GLIC), a prototypic pLGIC. The neurosteroid-based reagents photolabeled two sites per GLIC subunit, both within the transmembrane domain; one site was an intrasubunit site, and the other was located in the interface between subunits. By using reagents with photoreactive groups positioned throughout the neurosteroid backbone, we precisely map the orientation of neurosteroid binding within each site. Amino acid substitutions introduced at either site altered neurosteroid modulation of GLIC channel activity, demonstrating the functional role of both sites. These results provide a detailed molecular model of multisite neurosteroid modulation of GLIC, which may be applicable to other mammalian pLGICs. |
citation_standardnr | 6174196 |
datenlieferant | ipn_articles |
feed_id | 43 |
feed_publisher | The American Society for Biochemistry and Molecular Biology (ASBMB) |
feed_publisher_url | http://www.asbmb.org/ |
insertion_date | 2018-02-24 |
journaleissn | 1083-351X |
journalissn | 0021-9258 |
publikationsjahr_anzeige | 2018 |
publikationsjahr_facette | 2018 |
publikationsjahr_intervall | 7984:2015-2019 |
publikationsjahr_sort | 2018 |
publisher | The American Society for Biochemistry and Molecular Biology (ASBMB) |
quelle | Journal of Biological Chemistry |
relation | http://feedproxy.google.com/~r/jbc/SUcv/~3/vyrn0SnlPH0/3013.short |
search_space | articles |
shingle_author_1 | Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers |
shingle_author_2 | Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers |
shingle_author_3 | Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers |
shingle_author_4 | Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers |
shingle_catch_all_1 | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] Neurosteroids are endogenous sterols that potentiate or inhibit pentameric ligand-gated ion channels (pLGICs) and can be effective anesthetics, analgesics, or anti-epileptic drugs. The complex effects of neurosteroids on pLGICs suggest the presence of multiple binding sites in these receptors. Here, using a series of novel neurosteroid-photolabeling reagents combined with top-down and middle-down mass spectrometry, we have determined the stoichiometry, sites, and orientation of binding for 3α,5α-pregnane neurosteroids in the Gloeobacter ligand-gated ion channel (GLIC), a prototypic pLGIC. The neurosteroid-based reagents photolabeled two sites per GLIC subunit, both within the transmembrane domain; one site was an intrasubunit site, and the other was located in the interface between subunits. By using reagents with photoreactive groups positioned throughout the neurosteroid backbone, we precisely map the orientation of neurosteroid binding within each site. Amino acid substitutions introduced at either site altered neurosteroid modulation of GLIC channel activity, demonstrating the functional role of both sites. These results provide a detailed molecular model of multisite neurosteroid modulation of GLIC, which may be applicable to other mammalian pLGICs. Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers The American Society for Biochemistry and Molecular Biology (ASBMB) 0021-9258 00219258 1083-351X 1083351X |
shingle_catch_all_2 | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] Neurosteroids are endogenous sterols that potentiate or inhibit pentameric ligand-gated ion channels (pLGICs) and can be effective anesthetics, analgesics, or anti-epileptic drugs. The complex effects of neurosteroids on pLGICs suggest the presence of multiple binding sites in these receptors. Here, using a series of novel neurosteroid-photolabeling reagents combined with top-down and middle-down mass spectrometry, we have determined the stoichiometry, sites, and orientation of binding for 3α,5α-pregnane neurosteroids in the Gloeobacter ligand-gated ion channel (GLIC), a prototypic pLGIC. The neurosteroid-based reagents photolabeled two sites per GLIC subunit, both within the transmembrane domain; one site was an intrasubunit site, and the other was located in the interface between subunits. By using reagents with photoreactive groups positioned throughout the neurosteroid backbone, we precisely map the orientation of neurosteroid binding within each site. Amino acid substitutions introduced at either site altered neurosteroid modulation of GLIC channel activity, demonstrating the functional role of both sites. These results provide a detailed molecular model of multisite neurosteroid modulation of GLIC, which may be applicable to other mammalian pLGICs. Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers The American Society for Biochemistry and Molecular Biology (ASBMB) 0021-9258 00219258 1083-351X 1083351X |
shingle_catch_all_3 | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] Neurosteroids are endogenous sterols that potentiate or inhibit pentameric ligand-gated ion channels (pLGICs) and can be effective anesthetics, analgesics, or anti-epileptic drugs. The complex effects of neurosteroids on pLGICs suggest the presence of multiple binding sites in these receptors. Here, using a series of novel neurosteroid-photolabeling reagents combined with top-down and middle-down mass spectrometry, we have determined the stoichiometry, sites, and orientation of binding for 3α,5α-pregnane neurosteroids in the Gloeobacter ligand-gated ion channel (GLIC), a prototypic pLGIC. The neurosteroid-based reagents photolabeled two sites per GLIC subunit, both within the transmembrane domain; one site was an intrasubunit site, and the other was located in the interface between subunits. By using reagents with photoreactive groups positioned throughout the neurosteroid backbone, we precisely map the orientation of neurosteroid binding within each site. Amino acid substitutions introduced at either site altered neurosteroid modulation of GLIC channel activity, demonstrating the functional role of both sites. These results provide a detailed molecular model of multisite neurosteroid modulation of GLIC, which may be applicable to other mammalian pLGICs. Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers The American Society for Biochemistry and Molecular Biology (ASBMB) 0021-9258 00219258 1083-351X 1083351X |
shingle_catch_all_4 | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] Neurosteroids are endogenous sterols that potentiate or inhibit pentameric ligand-gated ion channels (pLGICs) and can be effective anesthetics, analgesics, or anti-epileptic drugs. The complex effects of neurosteroids on pLGICs suggest the presence of multiple binding sites in these receptors. Here, using a series of novel neurosteroid-photolabeling reagents combined with top-down and middle-down mass spectrometry, we have determined the stoichiometry, sites, and orientation of binding for 3α,5α-pregnane neurosteroids in the Gloeobacter ligand-gated ion channel (GLIC), a prototypic pLGIC. The neurosteroid-based reagents photolabeled two sites per GLIC subunit, both within the transmembrane domain; one site was an intrasubunit site, and the other was located in the interface between subunits. By using reagents with photoreactive groups positioned throughout the neurosteroid backbone, we precisely map the orientation of neurosteroid binding within each site. Amino acid substitutions introduced at either site altered neurosteroid modulation of GLIC channel activity, demonstrating the functional role of both sites. These results provide a detailed molecular model of multisite neurosteroid modulation of GLIC, which may be applicable to other mammalian pLGICs. Wayland W. L. Cheng, Zi-Wei Chen, John R. Bracamontes, Melissa M. Budelier, Kathiresan Krishnan, Daniel J. Shin, Cunde Wang, Xin Jiang, Douglas F. Covey, Gustav Akk, Alex S. Evers The American Society for Biochemistry and Molecular Biology (ASBMB) 0021-9258 00219258 1083-351X 1083351X |
shingle_title_1 | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] |
shingle_title_2 | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] |
shingle_title_3 | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] |
shingle_title_4 | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] |
timestamp | 2025-06-30T23:33:00.026Z |
titel | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] |
titel_suche | Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC [Neurobiology] |
topic | W V |
uid | ipn_articles_6174196 |