Cysteine 893 is a target of regulatory thiol modifications of GluA1 AMPA receptors.
Recent studies indicate that glutamatergic signaling involves, and is regulated by, thiol modifying and redox-active compounds. In this study, we examined the role of a reactive cysteine residue, Cys-893, in the cytosolic C-terminal tail of GluA1 AMPA receptor as a potential regulatory target. Elimination of the thiol function by substitution of serine for Cys-893 led to increased steady-state expression level and strongly reduced interaction with SAP97, a major cytosolic interaction partner of GluA1 C-terminus. Moreover, we found that of the three cysteine residues in GluA1 C-terminal tail, Cys-893 is the predominant target for S-nitrosylation induced by exogenous nitric oxide donors in cultured cells and lysates. Co-precipitation experiments provided evidence for native association of SAP97 with neuronal nitric oxide synthase (nNOS) and for the potential coupling of Ca2+-permeable GluA1 receptors with nNOS via SAP97. Our results show that Cys-893 can serve as a molecular target for regulatory thiol modifications of GluA1 receptors, including the effects of nitric oxide.
Department of Biosciences, Division of Biochemistry and Biotechnology, University of Helsinki, Helsinki, Finland.
Lotta
L
von Ossowski
Turku Centre for Biotechnology, Åbo Akademi University and University of Turku, Turku, Finland.|A.I. Virtanen Institute, University of Eastern Finland, Kuopio, Finland.
Li-Li
LL
Li
Department of Biosciences, Division of Biochemistry and Biotechnology, University of Helsinki, Helsinki, Finland.
Tommi
T
Möykkynen
Department of Biosciences, Division of Biochemistry and Biotechnology, University of Helsinki, Helsinki, Finland.
Sarah K
SK
Coleman
Turku Centre for Biotechnology, Åbo Akademi University and University of Turku, Turku, Finland.
Michael J
MJ
Courtney
Department of Biosciences, Division of Biochemistry and Biotechnology, University of Helsinki, Helsinki, Finland.
Kari
http://orcid.org/0000-0001-5759-5533
K
Keinänen
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D019943:Amino Acid Substitution; D020134:Catalytic Domain; D003545:Cysteine; D000074288:Discs Large Homolog 1 Protein; D057809:HEK293 Cells / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism; D018091:Receptors, AMPA / Q000737:chemistry / Q000378:metabolism*; D012694:Serine
D048868:Adaptor Proteins, Signal Transducing; D003545:Cysteine; D000074288:Discs Large Homolog 1 Protein; D008565:Membrane Proteins; D052248:Nitric Oxide Synthase Type I; D018091:Receptors, AMPA; D012694:Serine; C513314:DLG1 protein, human