Reboud A M, Buisson M, Arpin M, Reboud J P
Biochim Biophys Acta. 1977 Feb 16;474(4):578-87. doi: 10.1016/0005-2787(77)90077-6.
40- and 60-S ribosomal subunits and 80-S ribosomes from rat liver were highly labelled by reductive methylation using formaldehyde and sodium boro-[3H] hydride, under conditions which did not decrease their activity in poly-U-directed polyphenylalanine synthesis. Dissociation of the monosomes, subunits dimers, and polysomes into free subunits was observed after methylation. Free proteins labelled after extraction from the ribosomal subunits incorporated 7 times more radioactivity than when labelled in the subunits. Proteins extracted from methylated subunits and ribosomes were analyzed by two-dimensional gel electrophoresis, and the radioactivity of each protein was compared to that of the same free protein. A classification of the proteins was established according to their accessibility to the reagents in the subunits and the ribosomes.
在不降低其在多聚尿苷指导的聚苯丙氨酸合成中活性的条件下，使用甲醛和硼氢化钠-[3H]对大鼠肝脏的40-S和60-S核糖体亚基以及80-S核糖体进行还原甲基化，可使其高度标记。甲基化后观察到单体、亚基二聚体和多聚体解离为游离亚基。从核糖体亚基中提取后标记的游离蛋白质比在亚基中标记时掺入的放射性多7倍。通过二维凝胶电泳分析从甲基化亚基和核糖体中提取的蛋白质，并将每种蛋白质的放射性与相同游离蛋白质的放射性进行比较。根据蛋白质在亚基和核糖体中对试剂的可及性建立了蛋白质分类。
