Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Structure. 2012 Jun 6;20(6):967-76. doi: 10.1016/j.str.2012.04.010.
Structural studies of human G protein-coupled receptors (GPCRs) have recently been accelerated through the use of a fusion partner that was inserted into the third intracellular loop. Using chimeras of the human β(2)-adrenergic and human A(2A) adenosine receptors, we present the methodology and data for the initial selection of an expanded set of fusion partners for crystallizing GPCRs. In particular, use of the thermostabilized apocytochrome b(562)RIL as a fusion partner displays certain advantages over previously utilized fusion proteins, resulting in a significant improvement in stability and structure of GPCR-fusion constructs.
通过使用插入到第三细胞内环的融合伴侣,最近加速了对人类 G 蛋白偶联受体(GPCR)的结构研究。利用人β(2)-肾上腺素能受体和人 A(2A)腺苷受体嵌合体,我们提出了用于初始选择一组扩展的融合伴侣以结晶 GPCR 的方法和数据。特别是,使用热稳定化脱细胞色素 b(562)RIL 作为融合伴侣显示出相对于先前使用的融合蛋白的某些优势,导致 GPCR-融合构建体的稳定性和结构得到显著改善。