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N-terminal T4 lysozyme fusion facilitates crystallization of a G protein coupled receptor.

ABSTRACT: A highly crystallizable T4 lysozyme (T4L) was fused to the N-terminus of the ?(2) adrenergic receptor (?(2)AR), a G-protein coupled receptor (GPCR) for catecholamines. We demonstrate that the N-terminal fused T4L is sufficiently rigid relative to the receptor to facilitate crystallogenesis without thermostabilizing mutations or the use of a stabilizing antibody, G protein, or protein fused to the 3rd intracellular loop. This approach adds to the protein engineering strategies that enable crystallographic studies of GPCRs alone or in complex with a signaling partner.

SUBMITTER: Zou Y

PROVIDER: S-EPMC3464249 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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N-terminal T4 lysozyme fusion facilitates crystallization of a G protein coupled receptor.

Zou Yaozhong Y Weis William I WI Kobilka Brian K BK

PloS one 20121004 10

A highly crystallizable T4 lysozyme (T4L) was fused to the N-terminus of the β(2) adrenergic receptor (β(2)AR), a G-protein coupled receptor (GPCR) for catecholamines. We demonstrate that the N-terminal fused T4L is sufficiently rigid relative to the receptor to facilitate crystallogenesis without thermostabilizing mutations or the use of a stabilizing antibody, G protein, or protein fused to the 3rd intracellular loop. This approach adds to the protein engineering strategies that enable crystal ...[more]

PMID: 23056231

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