n-terminal t4 lysozyme fusion facilitates crystallization of a g protein coupled receptor氨基端t4溶菌酶融合促进结晶的g蛋白耦合的受体.pdfVIP

N-Terminal T4 Lysozyme Fusion Facilitates Crystallization of a G Protein Coupled Receptor 1 1,2 1 Yaozhong Zou , William I. Weis , Brian K. Kobilka * 1 Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California, United States of America, 2 Department of Structural Biology, Stanford University School of Medicine, Stanford, California, United States of America Abstract A highly crystallizable T4 lysozyme (T4L) was fused to the N-terminus of the b adrenergic receptor (b AR), a G-protein 2 2 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. Citation: Zou Y, Weis WI, Kobilka BK (2012) N-Terminal T4 Lysozyme Fusion Facilitates Crystallization of a G Protein Coupled Receptor. PLoS ONE 7(10): e46039. doi:10.1371/journal.pone.0046039 Editor: Roland Seifert, Medical School of Hannover, United States of America Received May 31, 2012; Accepted August 28, 2012; Published October 4, 2012 Copyright: 2012 Zou et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: The authors acknowledge support f