2J4Y
Crystal structure of a rhodopsin stabilizing mutant expressed in mammalian cells
Summary for 2J4Y
| Entry DOI | 10.2210/pdb2j4y/pdb |
| Related | 1EDS 1EDV 1EDW 1EDX 1F88 1FDF 1GZM 1HZX 1JFP 1L9H 1LN6 1N3M 1NZS 1OV0 1OV1 1U19 1VQX |
| Descriptor | RHODOPSIN, RETINAL, 2-acetamido-2-deoxy-beta-D-glucopyranose (3 entities in total) |
| Functional Keywords | chromophore, lipoprotein, glycoprotein, sensory transduction, photoreceptor protein, integral membrane protein, g-protein coupled receptor, vision, membrane, receptor, palmitate, transducer, retinal protein, phosphorylation, signaling protein, photoreceptor, transmembrane, visual pigment |
| Biological source | BOS TAURUS (CATTLE) |
| Total number of polymer chains | 2 |
| Total formula weight | 79080.46 |
| Authors | Standfuss, J.,Xie, G.,Edwards, P.C.,Burghammer, M.,Oprian, D.D.,Schertler, G.F.X. (deposition date: 2006-09-07, release date: 2007-09-25, Last modification date: 2024-10-23) |
| Primary citation | Standfuss, J.,Xie, G.,Edwards, P.C.,Burghammer, M.,Oprian, D.D.,Schertler, G.F.X. Crystal Structure of a Thermally Stable Rhodopsin Mutant. J.Mol.Biol., 372:1179-, 2007 Cited by PubMed Abstract: 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. PubMed: 17825322DOI: 10.1016/J.JMB.2007.03.007 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (3.4 Å) |
Structure validation
Download full validation report
