2J4Y

Crystal structure of a rhodopsin stabilizing mutant expressed in mammalian cells

2J4Y の概要

• エントリーDOI: 10.2210/pdb2j4y/pdb
• 関連するPDBエントリー: 1EDS 1EDV 1EDW 1EDX 1F88 1FDF 1GZM 1HZX 1JFP 1L9H 1LN6 1N3M 1NZS 1OV0 1OV1 1U19 1VQX
• 分子名称: RHODOPSIN, RETINAL, 2-acetamido-2-deoxy-beta-D-glucopyranose (3 entities in total)
• 機能のキーワード: 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
• 由来する生物種: BOS TAURUS (CATTLE)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 79080.46

構造登録者

Standfuss, J.,Xie, G.,Edwards, P.C.,Burghammer, M.,Oprian, D.D.,Schertler, G.F.X. (登録日: 2006-09-07, 公開日: 2007-09-25, 最終更新日: 2024-10-23)

主引用文献

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

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: 17825322
DOI: 10.1016/J.JMB.2007.03.007

実験手法

X-RAY DIFFRACTION (3.4 Å)