UREA GRADIENT SDS-PAGE - A USEFUL TOOL IN THE INVESTIGATION OF SIGNAL TRANSDUCING G-PROTEINS

被引:21
作者
CODINA, J
GRENET, D
CHANG, KJ
BIRNBAUMER, L
机构
[1] BAYLOR UNIV,DEPT CELL BIOL,HOUSTON,TX 77030
[2] BAYLOR UNIV,DEPT MOLEC PHYSIOL & BIOPHYS,HOUSTON,TX 77030
[3] BURROUGHS WELLCOME CO,WELLCOME RES LABS,DIV CELL BIOL,RES TRIANGLE PK,NC 27709
[4] BAYLOR UNIV,DIV NEUROSCI,HOUSTON,TX 77030
来源
JOURNAL OF RECEPTOR RESEARCH | 1991年 / 11卷 / 1-4期
关键词
D O I
10.3109/10799899109066429
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We describe an updated and improved protocol to perform urea gradient/SDS-PAGE in which proteins are electrophoresed through 9% polyacrylamide gel slabs in the presence of a linear 4 M to 8 M gradient of urea using Laemmli's separation buffers. We provide examples of this technique to separate PTX labeled G protein alpha subunits, as well as unlabeled alpha and beta subunits of G proteins. Applications of the technique are exemplified in which (1) the chromatographic separations of G proteins in DEAE-Toyopearl and MonoQ columns are compared, (2) the complexity of PTX substrates expressed in human erythrocytes, bovine brain, dog ventricle, FRTL-5 cells, HIT cells, GH4C1 cells and RIN cells are compared, and (3) the polypeptide composition of G protein beta-gamma subunits, as expressed in several tissues and found in three distinct G proteins from a single cell population, are analyzed.
引用
收藏
页码:587 / 601
页数:15
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