Urea gradient/SDS-PAGE; a useful tool in the investigation of signal transducing G proteins
J Codina, D Grenet, KJ Chang… - Journal of receptor …, 1991 - Taylor & Francis
J Codina, D Grenet, KJ Chang, L Birnbaumer
Journal of receptor research, 1991•Taylor & FrancisWe 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 α subunits, as well as
unlabeled α and β subunits of G proteins. Applications of the technique are exemplified in
which (1) the chromatographic separations of G proteins in DEAE-Toyopearl and MonoQ …
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 α subunits, as well as
unlabeled α and β subunits of G proteins. Applications of the technique are exemplified in
which (1) the chromatographic separations of G proteins in DEAE-Toyopearl and MonoQ …
Abstract
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 α subunits, as well as unlabeled α and β 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 βγ subunits, as expressed in several tissues and found in three distinct G proteins from a single cell population, are analyzed.
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