Nikolaeva A I, Logunova N N
Biokhimiia. 1983 Jun;48(6):1042-51.
The properties of cytoplasmic mRNP particles from spleen lymphocytes (sedimentation coefficients and buoyant density) and the kinetics of incorporation of impulse amino acid label at definite labelling periods (5, 15, 30, 60 sec) were studied. After 5 or 15 sec labelling periods the labelled amino acid was detected in the polysomes (sedimentation coefficient 130-240S, buoyant density 1.55 g/cm3) and in small-sized particles (sedimentation coefficient 80S and less, buoyant density 1.34, 1.50 and 1.60 g/cm3). After 30 sec of labelling the label is incorporated into the medium-sized particles (sedimentation coefficient 80-120 S, buoyant density 1.47 g/cm3); no increase of the label incorporation into the small-sized particles is observed. After 60 sec the label content in all the three sucrose gradient zones is increased. The observed succession of the amino acid label incorporation first into the small-sized particles and then into the large-sized ones having a different buoyant density and the data from impulse uridyl labelling suggest that in the lymphocytes the translation occurs already in the complexes of informosomes with single ribosomes and continues in the course of their conversion to polysomes. The short-time labelling of the cells with an impulse amino acid label allows to determine the time of the polypeptide chain translation from the time of polysome saturation with the label.
研究了脾淋巴细胞胞质mRNA颗粒的性质(沉降系数和浮力密度)以及在特定标记时间段(5、15、30、60秒)脉冲氨基酸标记的掺入动力学。在5秒或15秒的标记时间段后,在多核糖体(沉降系数130 - 240S,浮力密度1.55 g/cm³)和小颗粒(沉降系数80S及以下,浮力密度1.34、1.50和1.60 g/cm³)中检测到标记氨基酸。标记30秒后,标记物掺入中等大小颗粒(沉降系数80 - 120S,浮力密度1.47 g/cm³);未观察到小颗粒中标记掺入量增加。60秒后,所有三个蔗糖梯度区的标记物含量均增加。观察到的氨基酸标记先掺入小颗粒,然后掺入具有不同浮力密度的大颗粒的顺序以及脉冲尿苷标记的数据表明,在淋巴细胞中,翻译已经在信息体与单个核糖体的复合物中发生,并在它们转化为多核糖体的过程中继续进行。用脉冲氨基酸标记对细胞进行短时间标记,可以从多核糖体被标记饱和的时间来确定多肽链翻译的时间。