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克隆大鼠神经胶质细胞单层培养物中脑特异性蛋白质合成的调控

Regulation of synthesis of a brain-specific protein in monolayer cultures of clonal rat glial cells.

作者信息

Labourdette G, Mahony J B, Brown I R, Marks A

出版信息

Eur J Biochem. 1977 Dec;81(3):591-7. doi: 10.1111/j.1432-1033.1977.tb11986.x.

Abstract

C6 cells were grown in monolayer culture under conditions permitting continued exponential cell division after attainment of a density at which extensive intercellular contacts were formed. An increase in the relative synthesis of S100 protein coincided with the time of formation of extensive intercellular contacts and preceded the onset of the stationary phase of growth by three generations. These observations suggested that the induction of S100 protein synthesis was mediated by cell contact and not by an arrest of cellular growth. The mechanism of this induction was first studied in a homologous non-initiating cell-free protein-synthesizing system from C6 cells, using fixed amounts of free amino acids or fully charged rat liver aminoacyl-tRNA as a source of precursors for protein synthesis. Real synthesis of total soluble proteins decreased as the cells progressed from logarithmic to stationary growth while synthesis of S100 protein increased during this period. The capacity of poly(A)+ RNA from logarithmic and stationary cultures to direct the synthesis of S100 protein was estimated in a cell-free protein-synthesizing system derived from wheat embryos. Increased synthesis of S100 protein in stationary cultures was directly correlated with an increase in translatable S100 protein mRNA.

摘要

C6细胞在单层培养条件下生长,当达到形成广泛细胞间接触的密度后,允许细胞持续进行指数分裂。S100蛋白相对合成量的增加与广泛细胞间接触的形成时间一致,且在生长稳定期开始前三代就已出现。这些观察结果表明,S100蛋白合成的诱导是由细胞接触介导的,而非细胞生长停滞所致。这种诱导机制首先在来自C6细胞的同源非起始无细胞蛋白质合成系统中进行研究,使用固定量的游离氨基酸或完全负载的大鼠肝脏氨酰-tRNA作为蛋白质合成的前体来源。随着细胞从对数生长期进入稳定生长期,总可溶性蛋白质的实际合成量下降,而在此期间S100蛋白的合成量增加。在源自小麦胚的无细胞蛋白质合成系统中,评估了对数期和稳定期培养物中聚腺苷酸加尾RNA(poly(A)+ RNA)指导S100蛋白合成的能力。稳定期培养物中S100蛋白合成的增加与可翻译的S100蛋白mRNA的增加直接相关。

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