Redox Regulation of Signaling Complex between Caveolin-1 and Neuronal Calcium Sensor Recoverin

• Published in: Biomolecules (Basel, Switzerland) Vol. 12; no. 11; p. 1698
• Main Authors: Vladimirov, Vasiliy I, Shchannikova, Margarita P, Baldin, Alexey V, Kazakov, Alexey S, Shevelyova, Marina P, Nazipova, Aliya A, Baksheeva, Viktoriia E, Nemashkalova, Ekaterina L, Frolova, Anastasia S, Tikhomirova, Natalia K, Philippov, Pavel P, Zamyatnin, Andrey A, Permyakov, Sergei E, Zinchenko, Dmitry V, Zernii, Evgeni Yu
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01.11.2022; MDPI
• Subjects: Affinity; Amino acids; Apoptosis; Calcium (intracellular); Calcium signalling; Calcium-binding proteins; Caveolin; Caveolin-1; Caveolins; Cellular signal transduction; Cholesterol; Cytoplasm; Dimerization; Enzymes; Homeostasis; Intracellular; Kinases; Light; Light effects; Lipid rafts; Localization; Mimicry; Molecular biology; Monoclonal antibodies; Mutants; Oxidation; Oxidation-reduction reaction; Oxidative stress; Phosphorylation; photoreceptor; Photoreceptors; Phototransduction; Physiological aspects; Protein folding; Proteins; Reagents; Recoverin; Retina; Rhodopsin kinase; Signal transduction; Zinc; Russia; United States > US
• ISSN: 2218-273X; 2218-273X
• DOI: 10.3390/biom12111698

Caveolin-1 is a cholesterol-binding scaffold protein, which is localized in detergent-resistant membrane (DRM) rafts and interacts with components of signal transduction systems, including visual cascade. Among these components are neuronal calcium sensors (NCSs), some of which are redox-sensitive proteins that respond to calcium signals by modulating the activity of multiple intracellular targets. Here, we report that the formation of the caveolin-1 complex with recoverin, a photoreceptor NCS serving as the membrane-binding regulator of rhodopsin kinase (GRK1), is a redox-dependent process. Biochemical and biophysical in vitro experiments revealed a two-fold decreased affinity of recoverin to caveolin-1 mutant Y14E mimicking its oxidative stress-induced phosphorylation of the scaffold protein. At the same time, wild-type caveolin-1 demonstrated a 5–10-fold increased affinity to disulfide dimer of recoverin (dRec) or its thiol oxidation mimicking the C39D mutant. The formation of dRec in vitro was not affected by caveolin-1 but was significantly potentiated by zinc, the well-known mediator of redox homeostasis. In the MDCK cell model, oxidative stress indeed triggered Y14 phosphorylation of caveolin-1 and disulfide dimerization of recoverin. Notably, oxidative conditions promoted the accumulation of phosphorylated caveolin-1 in the plasma membrane and the recruitment of recoverin to the same sites. Co-localization of these proteins was preserved upon depletion of intracellular calcium, i.e., under conditions reducing membrane affinity of recoverin but favoring its interaction with caveolin-1. Taken together, these data suggest redox regulation of the signaling complex between recoverin and caveolin-1. During oxidative stress, the high-affinity interaction of thiol-oxidized recoverin with caveolin-1/DRMs may disturb the light-induced translocation of the former within photoreceptors and affect rhodopsin desensitization.