Single-Molecule Imaging of a Fluorescent Unnatural Amino Acid Incorporated Into Nicotinic Receptors
Single-Molecule Imaging of a Fluorescent Unnatural Amino Acid Incorporated Into Nicotinic Receptors
复制标题
DOI：
10.1016/j.bpj.2008.09.034
复制
发表时间：
2009-01-07
影响因子：
3.4
通讯作者：
Lester, Henry A.
中科院分区：
文献类型：
--
作者：
Pantoja, Rigo;Rodriguez, Erik A.;Lester, Henry A.
我们报告的第一个，据我们所知，成功检测到的荧光非天然氨基酸（fUAA），赖氨酸（BODIPYFL），纳入膜蛋白（肌肉烟碱乙酰胆碱受体，nAChR）在活细胞。用移码抑制tRNA注射爪蟾卵母细胞，用Lys（BODIPYFL）和nAChR（α/β 19'GGGU/γ/δ）mRNA氨基酰化。我们使用时间分辨全内反射荧光显微镜测量了表达nAChR β 19'Lys（BODIPYFL）的卵母细胞的荧光。在受体密度相对较低的条件下（
We report on the first, to our knowledge, successful detection of a fluorescent unnatural amino acid (fUAA), Lys(BODIPYFL), incorporated into a membrane protein (the muscle nicotinic acetylcholine receptor, nAChR) in a living cell. Xenopus oocytes were injected with a frameshift-suppressor tRNA, amino-acylated with Lys(BODIPYFL) and nAChR (alpha/beta 19'GGGU/gamma/delta) mRNAs. We measured fluorescence from oocytes expressing nAChR beta 19'Lys(BODIPYFL), using time-resolved total internal reflection fluorescence microscopy. Under conditions of relatively low receptor density (