PURIFICATION OF EUKARYOTIC INITIATION FACTOR-EIF-2, FACTOR-EIF-2B AND FACTOR-EIF-2-ALPHA-KINASE FROM BOVINE LIVER

被引:3
|
作者
FELDHOFF, RC [1 ]
KARINCH, AM [1 ]
KIMBALL, SR [1 ]
JEFFERSON, LS [1 ]
机构
[1] PENN STATE UNIV,MILTON S HERSHEY MED CTR,COLL MED,DEPT CELLULAR & MOLEC PHYSIOL,HERSHEY,PA 17033
来源
PREPARATIVE BIOCHEMISTRY | 1993年 / 23卷 / 03期
关键词
D O I
10.1080/10826069308544562
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Eukaryotic initiation factors 2 and 2B (eIF-2; eIF-2B) are components of the rate-limiting step in the initiation of eukaryotic protein synthesis and are involved in the regulation of this process. When the alpha-subunit of eIF-2 is phosphorylated by an eIF-2alpha kinase, the phosphorylated eIF-2alpha (eIF-2alpha(P)) binds tightly to eIF-2B and prevents the recycling of eIF-2.GDP to eIF-2.GTP which is required for sustained initiation of protein synthesis. The minute quantities of these proteins which are present in rat liver and muscle cytosol along with hundreds of other proteins has hindered purification efforts, as well as structure:function and regulatory studies. Therefore, procedures were developed for the simultaneous purification of eIF-2, eIF-2B and eIF-2alpha kinase from kilogram quantities of fresh bovine liver. Briefly, the 0-45% ammonium sulfate precipitate of the 200,000 x g supernatant was solubilized and chromatographed on DEAE-cellulose, heparin-agarose, Mono Q, Mono S, and Superose columns. The availability of purified quantities of these factors will be useful for investigations of molecular mechanisms of action and antibody production.
引用
收藏
页码:363 / 374
页数:12
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