Recoverin as a redox-sensitive protein.
Recoverin is a member of the neuronal calcium sensor (NCS) family of EF-hand calcium binding proteins. In a visual cycle of photoreceptor cells, recoverin regulates activity of rhodopsin kinase in a Ca2+-dependent manner. Like all members of the NSC family, recoverin contains a conserved cysteine (Cys38) in nonfunctional EF-hand 1. This residue was shown to be critical for activation of target proteins in some members of the NCS family but not for interaction of recoverin with rhodopsin kinase. Spectrophotometric titration of Ca2+-loaded recoverin gave 7.6 for the pKa value of Cys38 thiol, suggesting partial deprotonation of the thiol in vivo conditions. An ability of recoverin to form a disulfide dimer and thiol-oxidized monomer under mild oxidizing conditions was found using SDS-PAGE in reducing and nonreducing conditions and Ellman's test. Both processes are reversible and modulated by Ca2+. Although formation of the disulfide dimer takes place only for Ca2+-loaded recoverin, accumulation of the oxidized monomer proceeds more effectively for apo-recoverin. The Ca2+ modulated susceptibility of the recoverin thiol to reversible oxidation may be of potential importance for functioning of recoverin in photoreceptor cells.
Permyakov
Sergei E
SE
Institute for Biological Instrumentation of the Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russia.
Nazipova
Aliya A
AA
Denesyuk
Alexander I
AI
Bakunts
Anush G
AG
Zinchenko
Dmitry V
DV
Lipkin
Valery M
VM
Uversky
Vladimir N
VN
Permyakov
Eugene A
EA
D002118:Calcium / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011487:Protein Conformation; D051603:Recoverin / Q000737:chemistry* / Q000378:metabolism; D055672:Static Electricity; D013438:Sulfhydryl Compounds / Q000737:chemistry
D004220:Disulfides; D013438:Sulfhydryl Compounds; D051603:Recoverin; D003545:Cysteine; D002118:Calcium