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德尔塔赖氨酸120,一种使核糖体蛋白L7/L12的核糖体结合结构域不稳定的突变。

Delta Lys120, a mutation which destabilizes the ribosome-binding domain of ribosomal protein L7/L12.

作者信息

Laughrea M, Higgins E

机构信息

Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, Montréal, Que., Canada.

出版信息

Biochem Cell Biol. 1990 May;68(5):832-8. doi: 10.1139/o90-123.

Abstract

Five-residue-long deletions centered on Ala63, Ala75, and Glu118 of ribosomal protein L7/L12 gave low mutant yields (5% or less) when the mutant genes were cloned in phage M13mp18 and controlled by the L10 promotor. Deletions of Glu118-Lys120 or Lys120 (the COOH-terminus of L7/L12) gave higher mutant yields, up to 50% with L7/L12 delta Lys120. L7/L12 delta Lys120 was not preferentially found in the S100 and not preferentially removed by LiCl washing, but was preferentially extracted from 70S ribosomes in the presence of 28-35% ethanol in 0.25-0.5 M NH4Cl. It follows that delta Lys120 destabilizes the ribosome-binding domain of ribosomal protein L7/L12 in an ethanol-containing solvent, which raises the question whether Lys120 is part of the ribosome-binding domain of L7/L12 during some step of protein synthesis or whether it is essential to preserve the conformation of the physiological ribosome-binding domain under structurally stressful conditions.

摘要

当将突变基因克隆到噬菌体M13mp18中并由L10启动子控制时,以核糖体蛋白L7/L12的Ala63、Ala75和Glu118为中心的五个残基长的缺失产生的突变体产量较低(5%或更低)。Glu118 - Lys120或Lys120(L7/L12的COOH末端)的缺失产生的突变体产量更高,L7/L12缺失Lys120时高达50%。L7/L12缺失Lys120在S100中未被优先发现,也未被LiCl洗涤优先去除,但在0.25 - 0.5 M NH4Cl中存在28 - 35%乙醇的情况下,它优先从70S核糖体中被提取出来。由此可见,缺失Lys120会在含乙醇的溶剂中使核糖体蛋白L7/L12的核糖体结合结构域不稳定,这就引发了一个问题,即Lys120在蛋白质合成的某些步骤中是否是L7/L12核糖体结合结构域的一部分,或者在结构压力条件下它对于维持生理核糖体结合结构域的构象是否至关重要。

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