TY - JOUR
T1 - Native mass spectrometry reveals the simultaneous binding of lipids and zinc to rhodopsin
AU - Norris, Carolanne E.
AU - Keener, James E.
AU - Perera, Suchithranga M.D.C.
AU - Weerasinghe, Nipuna
AU - Fried, Steven D.E.
AU - Resager, William C.
AU - Rohrbough, James G.
AU - Brown, Michael F.
AU - Marty, Michael T.
N1 - Funding Information:
The authors thank Deseree Reid for assistance in sample analysis. This work was funded by the Bisgrove Scholar Award from Science Foundation Arizona and the National Institute of General Medical Sciences and National Institutes of Health under Award Number R35 GM128624 to M.T.M. Additional funding was provided by National Science Foundation awards CHE 1904125 and MCB 11817862 to M.F.B. and National Institutes of Health awards EY012049 and EY02604 to M.F.B. Support of S.M.D.C. was by a Technology Research Initiative Fund predoctoral fellowship from the Arizona Board of Regents . S.D.E.F. was partially supported by a Goldwater research scholarship. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
Rhodopsin was purified by SMDCP, NW, and SDEF with supervision and funding from MFB. Rhodopsin was prepared for native MS and analyzed by CEN, JEK, and JGR. WCR contributed to early native MS method development. Data was processed, analyzed, and visualized by CEN and MTM using software developed by MTM. The manuscript was written by CEN and MTM with contributions from MFB and other coauthors. Funding and project supervision were contributed by MTM.
Publisher Copyright:
© 2020 Elsevier B.V.
PY - 2021/2
Y1 - 2021/2
N2 - Rhodopsin, a prototypical G-protein-coupled receptor, is responsible for scoptic vision at low-light levels. Although rhodopsin's photoactivation cascade is well understood, it remains unclear how lipid and zinc binding to the receptor are coupled. Using native mass spectrometry, we developed a novel data analysis strategy to deconvolve zinc and lipid bound to the proteoforms of rhodopsin and investigated the allosteric interaction between lipids and zinc binding. We discovered that phosphatidylcholine bound to rhodopsin with a greater affinity than phosphatidylserine or phosphatidylethanolamine, and that binding of all lipids was influenced by zinc but with different effects. In contrast, zinc binding was relatively unperturbed by lipids. Overall, these data reveal that lipid binding can be strongly and differentially influenced by metal ions.
AB - Rhodopsin, a prototypical G-protein-coupled receptor, is responsible for scoptic vision at low-light levels. Although rhodopsin's photoactivation cascade is well understood, it remains unclear how lipid and zinc binding to the receptor are coupled. Using native mass spectrometry, we developed a novel data analysis strategy to deconvolve zinc and lipid bound to the proteoforms of rhodopsin and investigated the allosteric interaction between lipids and zinc binding. We discovered that phosphatidylcholine bound to rhodopsin with a greater affinity than phosphatidylserine or phosphatidylethanolamine, and that binding of all lipids was influenced by zinc but with different effects. In contrast, zinc binding was relatively unperturbed by lipids. Overall, these data reveal that lipid binding can be strongly and differentially influenced by metal ions.
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U2 - 10.1016/j.ijms.2020.116477
DO - 10.1016/j.ijms.2020.116477
M3 - Article
AN - SCOPUS:85096837258
VL - 460
JO - International Journal of Mass Spectrometry and Ion Processes
JF - International Journal of Mass Spectrometry and Ion Processes
SN - 1387-3806
M1 - 116477
ER -