de Herdt E, Piot E, Wahba A, Slegers H
Eur J Biochem. 1985 Sep 16;151(3):455-60. doi: 10.1111/j.1432-1033.1985.tb09123.x.
Non-polysomal poly(A)-containing mRNP of A. salina cryptobiotic embryos is separated in mRNP active in protein synthesis and in repressed mRNP by sucrose gradient centrifugation. In the translationally active fraction the presence of eukaryotic initiation factor 2 (eIF2) is demonstrated by electroblotting of sodium dodecylsulphate/polyacrylamide gels on nitrocellulose and anti-eIF2 antibody detection. mRNP proteins with Mr of 40 000 and 42 000 are identified as the alpha and beta subunits of eIF2. The repressed mRNP is devoid of eIF2 and is associated with an inhibitor ribonucleoprotein composed of a small 85 +/- 2-nucleotide-long RNA and a protein with Mr of 64 000. The latter ribonucleoprotein is a potent inhibitor of the translationally active mRNP.
通过蔗糖梯度离心,将盐生杜氏藻隐生胚的非多聚核糖体含多聚腺苷酸的mRNA核糖核蛋白体(mRNP)分离为具有蛋白质合成活性的mRNP和受抑制的mRNP。在翻译活性组分中,通过十二烷基硫酸钠/聚丙烯酰胺凝胶在硝酸纤维素膜上的电印迹和抗真核起始因子2(eIF2)抗体检测,证明了真核起始因子2(eIF2)的存在。分子量为40000和42000的mRNP蛋白被鉴定为eIF2的α和β亚基。受抑制的mRNP不含eIF2,并且与一种抑制性核糖核蛋白相关,该抑制性核糖核蛋白由一个85±2个核苷酸长的小RNA和一个分子量为64000的蛋白质组成。后者的核糖核蛋白是翻译活性mRNP的有效抑制剂。