2j4y

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Crystal structure of a rhodopsin stabilizing mutant expressed in mammalian cellsCrystal structure of a rhodopsin stabilizing mutant expressed in mammalian cells

Structural highlights

2j4y is a 2 chain structure with sequence from Bos taurus. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:,
NonStd Res:
Related:1eds, 1edv, 1edw, 1edx, 1f88, 1fdf, 1gzm, 1hzx, 1jfp, 1l9h, 1ln6, 1n3m, 1nzs, 1ov0, 1ov1, 1u19, 1vqx
Resources:FirstGlance, OCA, RCSB, PDBsum

Evolutionary Conservation

Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.

Publication Abstract from PubMed

We determined the structure of the rhodopsin mutant N2C/D282C expressed in mammalian cells; the first structure of a recombinantly produced G protein-coupled receptor (GPCR). The mutant was designed to form a disulfide bond between the N terminus and loop E3, which allows handling of opsin in detergent solution and increases thermal stability of rhodopsin by 10 deg.C. It allowed us to crystallize a fully deglycosylated rhodopsin (N2C/N15D/D282C). N15 mutations are normally misfolding and cause retinitis pigmentosa in humans. Microcrystallographic techniques and a 5 microm X-ray beam were used to collect data along a single needle measuring 5 microm x 5 microm x 90 microm. The disulfide introduces only minor changes but fixes the N-terminal cap over the beta-sheet lid covering the ligand-binding site, a likely explanation for the increased stability. This work allows structural investigation of rhodopsin mutants and shows the problems encountered during structure determination of GPCRs and other mammalian membrane proteins.

Crystal structure of a thermally stable rhodopsin mutant.,Standfuss J, Xie G, Edwards PC, Burghammer M, Oprian DD, Schertler GF J Mol Biol. 2007 Oct 5;372(5):1179-88. Epub 2007 Mar 12. PMID:17825322[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

See Also

References

  1. Standfuss J, Xie G, Edwards PC, Burghammer M, Oprian DD, Schertler GF. Crystal structure of a thermally stable rhodopsin mutant. J Mol Biol. 2007 Oct 5;372(5):1179-88. Epub 2007 Mar 12. PMID:17825322 doi:10.1016/j.jmb.2007.03.007

2j4y, resolution 3.40Å

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