Diffusion of green fluorescent protein in the aqueous-phase lumen of endoplasmic reticulum
Diffusion of green fluorescent protein in the aqueous-phase lumen of endoplasmic reticulum
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DOI：
10.1016/s0006-3495(99)77438-2
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发表时间：
1999-05-01
影响因子：
3.4
通讯作者：
Verkman, AS
中科院分区：
文献类型：
--
作者：
Dayel, MJ;Hom, EFY;Verkman, AS
内质网(ER)是新合成蛋白质加工和质量控制的主要部分。用绿色荧光蛋白(GFP)作为非侵入性探针，通过瞬时转染编码GFP(S65T/F64L突变体)的C端KDEL保留序列和上游催乳素分泌序列的GFP，将GFP靶向于CHO细胞的ER腔。重复激光照射直径2微米的固定斑点，导致整个细胞内ER相关绿色荧光蛋白完全漂白，表明内质网管腔持续存在。剩余金额(
The endoplasmic reticulum (ER) is the major compartment for the processing and quality control of newly synthesized proteins. Green fluorescent protein (GFP) was used as a noninvasive probe to determine the Viscous properties of the aqueous lumen of the ER, GFP was targeted to the ER lumen of CHO cells by transient transfection with cDNA encoding GFP (S65T/F64L mutant) with a C-terminus KDEL retention sequence and upstream prolactin secretory sequence. Repeated laser illumination of a fixed 2-mu m diameter spot resulted in complete bleaching of ER-associated GFP throughout the cell, indicating a continuous ER lumen. A residual amount (