Retinitis pigmentosa mutants provide insight into the role of the N-terminal cap in rhodopsin folding, structure, and function

Chikwado A. Opefi, Kieron South, Christopher A. Reynolds, Steven O. Smith, Philip J. Reeves

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Autosomal dominant retinitis pigmentosa (ADRP) mutants (T4K, N15S, T17M, V20G, P23A/H/L, and Q28H) in the N-terminal cap of rhodopsin misfold when expressed in mammalian cells. To gain insight into the causes of misfolding and to define the contributions of specific residues to receptor stability and function, we evaluated the responses of these mutants to 11-cisretinal pharmacological chaperone rescue or disulfide bondmediated repair. Pharmacological rescue restored folding in all mutants, but the purified mutant pigments in all cases were thermo-unstable and exhibited abnormal photobleaching, metarhodopsin II decay, and G protein activation. As a complementary approach, we superimposed this panel of ADRP mutants onto a rhodopsin background containing a juxtaposed cysteine pair (N2C/D282C) that forms a disulfide bond. This approach restored folding in T4K, N15S, V20G, P23A, and Q28H but not T17M, P23H, or P23L. ADRP mutant pigments obtained by disulfide bond repair exhibited enhanced stability, and some also displayed markedly improved photobleaching and signal transduction properties. Our major conclusionisthat the N-terminal cap stabilizes opsin during biosynthesis and contributes to the dark-state stability of rhodopsin. Comparison of these two restorative approaches revealed that the correct position of the cap relative to the extracellular loops is also required for optimal photochemistry and efficient G protein activation.

Original languageEnglish
Pages (from-to)33912-33926
Number of pages15
JournalJournal of Biological Chemistry
Volume288
Issue number47
Early online date8 Oct 2013
DOIs
Publication statusPublished - 22 Nov 2013
Externally publishedYes

Bibliographical note

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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