Palmitoylation of bovine opsin and its cysteine mutants in COS cells.

Abstract

Previously, bovine rhodopsin has been shown to be palmitoylated at cysteine residues 322 and 323.

The Significance of G Protein-Coupled Receptor Crystallography for Drug Discovery

“…These results bolstered the idea that rhodopsin, and not bacteriorhodopsin, would serve as a model for all GPCRs, leading to its adoption for modeling helical arrangements within the transmembrane bundle of GPCRs (Baldwin, 1993(Baldwin, , 1994Baldwin et al, 1997;Unger et al, 1997). These structural results were further advanced by mutagenesis and biochemical studies allowing spacial assignment of post-translational modifications such as disulfide bonds (Karnik et al, 1988;Karnik and Khorana, 1990), palmitoylation (Ovchinnikov et al, 1988;Karnik et al, 1993), and phosphorylation (Palczewski et al, 1991;Ohguro et al, 1993Ohguro et al, , 1996Ohguro et al, , 1998. These biochemical studies together with specific mutations led to a clear demarcation of the border between loops and transmembrane regions (for review, see Menon et al, 2001;Sakmar et al, 2002;Filipek et al, 2003a;Hubbell et al, 2003;Palczewski, 2006).…”

Neurotensin receptor-1 inducible palmitoylation is required for efficient receptor-mediated mitogenic-signaling within structured membrane microdomains

“…To confirm and validate the specificity of glycosylated NTSR-1 species, we assessed N-and (GPCR) to be identified as a target for palmitoylation. 7 GPCR palmitoylation alters receptor conformation and thereby, regulates the interactions of the receptor with specific downstream effectors. 8 A palmitoylation/depalmitoylation cycle upon stimulation was observed for several GPCRs, such as the D1 dopamine, [9][10][11] β2-adrenergic 12 and α2A-adrenergic receptors.…”

G Protein-Coupled Receptor Rhodopsin: A Prospectus

“…The presence of these palmitoyl groups, attached enzymatically or spontaneously (46,95), and the loop between helix VII and VIII may affect rhodopsin regeneration (72,96). In COS cells, Cys 322 and Cys 323 mutants showed properties similar to those of wild-type rhodopsin (97). It has been proposed that this region of helix VIII and palmitoylation is the site of non-covalent binding of all-trans-retinal to opsin (98), in agreement with rhodopsin modeling (D.C. Teller, unpublished data).…”