Comparison of refolding activities between nanogel artificial chaperone and GroEL systems
Comparison of refolding activities between nanogel artificial chaperone and GroEL systems
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DOI：
10.1016/j.ijbiomac.2007.11.003
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发表时间：
2008-04-01
影响因子：
8.2
通讯作者：
Akiyoshi, Kazunari
中科院分区：
文献类型：
--
作者：
Asayama, Wakiko;Sawada, Shin-Ichi;Akiyoshi, Kazunari
比较了含胆固醇基团的普鲁兰(CHP)纳米凝胶和GroEL纳米凝胶对酸变性绿色荧光蛋白(GFP)的分子伴侣活性。在纳米凝胶或GroEL存在下，通过稀释酸变性GFP来实现变性GFP的复性。随着加入到稀释缓冲液中的CHP纳米凝胶或GroEL浓度的增加，GFP的荧光被日益抑制。抑制GFP荧光恢复的50%浓度分别为0.03 mU M(GroEL)和0.08 mU M(CHP纳米凝胶)。通过在GroEL(0.20µM)体系中加入三磷酸腺苷或在纳米凝胶(0.20µM)中加入甲基-对环糊精来恢复复性。在纳米凝胶-GFP体系中，约90%的强度在10分钟内恢复。在CHP纳米凝胶体系(36 S)中复性的半衰期(t(1/2))几乎与天然伴侣GroEL-Groes体系相当。(C)2007 Elsevier B.V.保留所有权利。
The chaperone-like activity of a nanogel of cholesteryl group-bearing pullulan (CHP) was compared with that of GroEL for refolding acid-denatured green fluorescent protein (GFP). The refolding of denatured GFP was carried out by dilution of acid-denatured GFP in the presence of the nanogel or GroEL. GFP fluorescence was increasingly repressed with increases in the concentration of the CHP nanogel or GroEL added to the dilution buffer. The concentrations of 50% inhibition of recovery of GFP fluorescence were 0.03 mu M (GroEL) and 0.08 mu M (CHP nanogel), respectively. The refolding was resumed by the addition of ATP into the GroEL (0.20 mu M) system or by the addition of methyl-p-cyclodextrin into the nanogel (0.20 mu M) system. In the nanogel-GFP system, about 90% of the intensity was recovered within 10 min. The half time (t(1/2)) for refolding in the CHP nanogel system (36 s) is almost equal to that of the natural chaperone GroEL-GroES system. (C) 2007 Elsevier B.V. All rights reserved.