4U15

M3-mT4L receptor bound to tiotropium

Summary for 4U15

• Entry DOI: 10.2210/pdb4u15/pdb
• Related: 4U14 4U16
• Descriptor: Muscarinic acetylcholine receptor M3,Lysozyme,Muscarinic acetylcholine receptor M3, (1R,2R,4S,5S,7S)-7-{[hydroxy(dithiophen-2-yl)acetyl]oxy}-9,9-dimethyl-3-oxa-9-azoniatricyclo[3.3.1.0~2,4~]nonane, HEXAETHYLENE GLYCOL, ... (6 entities in total)
• Functional Keywords: gpcr t4l stabillized crystallography, membrane protein, membrane protein-inhibitor complex, membrane protein/inhibitor
• Biological source: Rattus norvegicus (Rat); More
• Cellular location: Cell membrane; Multi-pass membrane protein: P08483
• Total number of polymer chains: 2
• Total formula weight: 98943.82

Authors

Thorsen, T.S.,Matt, R.,Weis, W.I.,Kobilka, B. (deposition date: 2014-07-15, release date: 2014-11-26, Last modification date: 2024-11-20)

Primary citation

Thorsen, T.S.,Matt, R.,Weis, W.I.,Kobilka, B.K.
Modified T4 Lysozyme Fusion Proteins Facilitate G Protein-Coupled Receptor Crystallogenesis.
Structure, 22:1657-1664, 2014

PubMed Abstract: G protein-coupled receptors (GPCRs) mediate the majority of cellular responses to hormones and neurotransmitters. Most GPCR crystal structures have been obtained using a fusion protein strategy where the flexible third intracellular loop is replaced by T4 lysozyme (T4L). However, wild-type T4L may not be ideally suited for all GPCRs because of its size and the inherent flexibility between the N- and C-terminal subdomains. Here we report two modified T4L variants, designed to address flexibility and size, that can be used to optimize crystal quality or promote alternative packing interactions. These variants were tested on the M3 muscarinic receptor (M3). The original M3-T4L fusion protein produced twinned crystals that yielded a 3.4 Å structure from a 70 crystal data set. We replaced T4L with the modified T4L variants. Both T4L variants yielded M3 muscarinic receptor crystals with alternate lattices that were not twinned, including one that was solved at 2.8 Å resolution.

PubMed: 25450769
DOI: 10.1016/j.str.2014.08.022

Experimental method

X-RAY DIFFRACTION (2.8 Å)