RENATURATION OF AEQUOREA GREEN-FLUORESCENT PROTEIN
RENATURATION OF AEQUOREA GREEN-FLUORESCENT PROTEIN
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DOI：
10.1016/0006-291x(81)91599-0
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发表时间：
1981-01-01
影响因子：
3.1
通讯作者：
WARD, WW
中科院分区：
文献类型：
--
作者：
BOKMAN, SH;WARD, WW
用盐酸胍或酸变性后，首次实现了水母绿色荧光蛋白（A-GFP）的复性。变性伴随着可见荧光的共同损失，吸收特性的改变，以及远紫外区圆二色性信号的大的负偏转。透析的胍变性样品在pH 8导致64%的复性（返回到天然吸收）和中和的酸变性样品恢复90%的天然吸收。复性的GFP是高度荧光的，并且在激发和发射光谱的形状方面与天然GFP不可区分。天然和变性蛋白质都表现出对胰蛋白酶水解的抗性，并且具有相同的宽pH和热稳定性特征，所有这些都表明完全复性。
Renaturation of Aequorea green-fluorescent protein (A-GFP) was achieved for the 1st time following denaturation in guanidine-HCl or acid. Denaturation was accompanied by the concerted loss of visible fluorescence, alteration of absorption characteristics, and large negative deflection of circular dichroism signal in the far UV. Dialysis of a guanidine-denatured sample at pH 8 resulted in 64% renaturation (return to native absorption) and neutralization of an acid-denatured sample restored 90% of the native absorption. Renatured GFP is highly fluorescent and indistinguishable from native GFP with respect to the shape of excitation and emission spectra. Both native and denatured proteins exhibit resistance to trypsin hydrolysis and have identically broad pH and heat stability profiles, all of which suggest full renaturation.