HIGH-RESOLUTION ONE-DIMENSIONAL POLYACRYLAMIDE-GEL ISOELECTRIC-FOCUSING OF VARIOUS FORMS OF ELONGATION FACTOR-II

被引:26
作者
REDPATH, NT
机构
[1] Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol, BS8 1TD, University Walk
关键词
D O I
10.1016/0003-2697(92)90115-N
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A system for analyzing covalent modifications of elongation factor-2 (eEF-2) by one-dimensional isoelectric focusing in slab polyacrylamide gels is described. Depending on the degree of phosphorylation, four species of eEF-2 could be resolved corresponding to the un-, mono-, bis-, and trisphosphorylated factor. Furthermore, the degree of ADP-ribosylation of the protein could also be assessed by this method. It was also shown that an acidic isoform of eEF-2 exists which appears not to be artifactual and that the relative level of this isoform appeared to vary between different cell types. By Western blotting the gels and using an antibody against eEF-2 it is possible to assess the state of phosphorylation of the factor in cells. © 1992.
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页码:340 / 343
页数:4
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