Fusion partner toolchest for the stabilization and crystallization of G protein-coupled receptors.

Fusion partner toolchest for the stabilization and crystallization of G protein-coupled receptors.
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DOI:
10.1016/j.str.2012.04.010
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发表时间:
2012-06-06
期刊:
影响因子:
5.7
通讯作者:
Stevens, Raymond C.
Stevens, Raymond C.
中科院分区:
生物学2区
文献类型:
--
作者:
Chun, Eugene;Thompson, Aaron A.;Liu, Wei;Roth, Christopher B.;Griffith, Mark T.;Katritch, Vsevolod;Kunken, Joshua;Xu, Fei;Cherezov, Vadim;Hanson, Michael A.;Stevens, Raymond C.

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最近,通过使用插入第三胞内环的T4溶菌酶融合伴侣,加速了对人G蛋白偶联受体(GPCR)的结构研究。使用人β2-肾上腺素能受体和人A2 A腺苷受体的嵌合体,我们提出了用于选择用于结晶GPCR的五种新融合伴侣的方法和数据。特别地,使用热稳定的脱辅基细胞色素b562 RIL作为融合伴侣显示出优于先前使用的T4溶菌酶的某些优点,导致GPCR融合构建体的稳定性和结构的显著改善。
Structural studies of human G protein-coupled receptors (GPCRs) have recently been accelerated through the use of the T4 lysozyme fusion partner that was inserted into the third intracellular loop. Using chimeras of the human β2-adrenergic and human A2A adenosine receptors, we present the methodology and data for the selection of five new fusion partners for crystallizing GPCRs. In particular, the use of the thermostabilized apocytochrome b562RIL as a fusion partner displays certain advantages over the previously utilized T4 lysozyme, resulting in a significant improvement in stability and structure in GPCR-fusion constructs.
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