Renaud J, Eggo M C, Waithe W I
Ann Immunol (Paris). 1977 Jul-Oct;128C(4-5):909-13.
Lymphocytes activated by phytohaemagglutinin (PHA) change from a resting state (GO) to an actively growing one (G1). To investigate the role of growth regulatory protein in this process we have determined the patterns of synthesis of acidic (non-histone, NHCP) and basic (histone) nuclear chromatin proteins, and of cytoplasmic proteins during PHA activation of human lymphocytes, Protein fractions isolated from a mixture of stimulated (pulse-labelled with 14C leucine) and unstimulated (labelled with 3H-leucine) lymphocytes were separated on SDS-polyacrylamide gel electrophoresis and the ratio of 14C/3H determined on 1 mm gel slices. Several peaks of increased synthesis of specific peptides are detected in all fractions within 12 h of activation. When activation is blocked by an amino acid deficiency, the pattern of label upon release of the inhibition by addition of the missing amino acid is different from that of uninhibited lymphcytes.
被植物血凝素(PHA)激活的淋巴细胞从静止状态(G0)转变为活跃生长状态(G1)。为了研究生长调节蛋白在此过程中的作用,我们测定了人淋巴细胞在PHA激活过程中酸性(非组蛋白,NHCP)和碱性(组蛋白)核染色质蛋白以及细胞质蛋白的合成模式。从受刺激(用14C亮氨酸脉冲标记)和未受刺激(用3H亮氨酸标记)的淋巴细胞混合物中分离出的蛋白质组分,在SDS-聚丙烯酰胺凝胶电泳上进行分离,并在1毫米凝胶切片上测定14C/3H的比值。在激活后的12小时内,所有组分中均检测到特定肽段合成增加的几个峰值。当激活因氨基酸缺乏而受阻时,通过添加缺失的氨基酸解除抑制后,标记模式与未受抑制的淋巴细胞不同。