FTIR analysis of GPCR activation using azido probes.

FTIR analysis of GPCR activation using azido probes.
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DOI:
10.1038/nchembio.167
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发表时间:
2009-06
影响因子:
14.8
通讯作者:
Sakmar TP
Sakmar TP
中科院分区:
生物学1区
文献类型:
--
作者:
Ye S;Huber T;Vogel R;Sakmar TP

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我们展示了使用琥珀密码子抑制技术将 IR 活性氨基酸对叠氮基-L-苯丙氨酸 (azidoF, 1) 定点掺入 G 蛋白偶联受体视紫红质。叠氮基团的反对称伸缩振动在清晰的光谱窗口中吸收约 2,100 cm−1 处,并且对其静电环境敏感。我们使用 FTIR 差异光谱来监测叠氮探针,并显示特定螺旋间网络的静电环境在受体激活过程中发生变化。
We demonstrate the site-directed incorporation of an IR-active amino acid, p-azido-l-phenylalanine (azidoF, 1), into the G protein–coupled receptor rhodopsin using amber codon suppression technology. The antisymmetric stretch vibration of the azido group absorbs at ~2,100 cm−1 in a clear spectral window and is sensitive to its electrostatic environment. We used FTIR difference spectroscopy to monitor the azido probe and show that the electrostatic environments of specific interhelical networks change during receptor activation.
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