Covalent and noncovalent interactions mediate metabotropic glutamate receptor mGlu5 dimerization

Carmelo Romano, Judith Kelvin Miller, Krzysztof Hyrc, Seta Dikranian, Steven Mennerick, Yutaka Takeuchi, Mark P. Goldberg, Karen L. O'Malley

Research output: Contribution to journalArticle

118 Scopus citations

Abstract

Some, perhaps all, G protein-coupled receptors form homo- or heterodimers. We have shown that metabotropic glutamate receptors are covalent dimers, held together by one or more disulfide bonds near the N terminus. Here we report how mutating cysteines in this region affect dimerization and function. Covalent dimerization is preserved when cysteines 57, 93, or 99 are mutated but lost with replacement at 129. Coimmunoprecipitation under nondenaturing conditions indicates that the C[129]S mutant receptor remains a dimer, via noncovalent interactions. Both C[93]S and C[129]S bind [3H]quisqualate, whereas binding to C[57]S or C[99]S mutants is absent or greatly attenuated. The C[93]S and C[129]S receptors have activity simiiar to wild-type when assayed by fura-2 imaging of intracellular calcium in human embryonic kidney cells or electrophysiologically in Xenopus laevis oocytes. In contrast, C[57]S or C[99]S are less active in both assays but do respond with higher glutamate concentrations in the oocyte assay. These results demonstrate that 1) covalent dimerization is not critical for mGlu5 binding or function; 2) mGlu5 remains a noncovalent dimer even in the absence of covalent dimerization; and 3) high-affinity binding requires Cys-57 and Cys-99.

Original languageEnglish
Pages (from-to)46-53
Number of pages8
JournalMolecular pharmacology
Volume59
Issue number1
DOIs
StatePublished - Jan 1 2001

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