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通过圆二色性测量洗涤剂对大肠杆菌转运RNA构象的影响。

Effects of detergents on the conformation of Escherichia coli tRNA as measured by circular dichroism.

作者信息

Okabe N, Serikawa M, Tomita K

出版信息

J Biochem. 1977 Apr;81(4):871-7. doi: 10.1093/oxfordjournals.jbchem.a131551.

Abstract

The effect of a cationic detergent, lauryl pyridiniumchloride (LPC), and an anionic one, sodium n-octylbenzenesulfonate (SOBS), on the conformation of unfractionated Escherichia coli tRNA was investigated at various molar ratios of detergent to tRNA (D/R) in the presence and absence of Mg2+ and Na+ ions by measuring the circular dichroism (CD) at 265 nm and 340 nm, which reflects conformational change involving base pairs and/or base stacking, and the disymmetry in the vicinity of 4-thiouridylate (4-TU), respectively. In the presence of Mg2+ and Na+ ions, the tRNA retains its native structure even in the presence of high molar ratios of detergent to tRNA (D/R congruent to 40 at 265 nm and D/R congruent to 20 at 340 nm). However, in the absence of these metal ions, the ellipticity at 340 nm was very sensitive to LPC concentration and decreased from 5,600 to nearly--1,600 at 25 degrees C with the increase of D/R ratios up to 20, and a similar decrease in the ellipticity at 340 nm was observed on thermal denaturation. This result suggests that the local environment involving the 4-TU region might be readily influenced by LPC, reflecting a large conformational change. However, no effect was observed in the case of the SOBS: tRNA system. On the other hand, secondary base pairing and/or base stacking structure was virtually invariant on adding both LPC and SOBS even at high D/R ratios in the absence of Mg2+ and Na+ ions.

摘要

通过测量265nm和340nm处的圆二色性(CD),研究了阳离子去污剂十二烷基氯化吡啶鎓(LPC)和阴离子去污剂正辛基苯磺酸钠(SOBS)在有无Mg2+和Na+离子存在的情况下,以不同的去污剂与tRNA摩尔比(D/R)对未分级的大肠杆菌tRNA构象的影响。265nm处的CD反映涉及碱基对和/或碱基堆积的构象变化,340nm处的CD分别反映4-硫尿苷酸(4-TU)附近的不对称性。在Mg2+和Na+离子存在的情况下,即使在去污剂与tRNA的摩尔比很高时(265nm处D/R等于40,340nm处D/R等于20),tRNA仍保持其天然结构。然而,在没有这些金属离子的情况下,340nm处的椭圆率对LPC浓度非常敏感,在25℃下,随着D/R比值增加到20,椭圆率从5600下降到近-1600,并且在热变性时也观察到340nm处椭圆率有类似的下降。这一结果表明,涉及4-TU区域的局部环境可能很容易受到LPC的影响,反映出较大的构象变化。然而,在SOBS:tRNA系统中未观察到影响。另一方面,即使在没有Mg2+和Na+离子的情况下,添加LPC和SOBS时,即使在高D/R比值下,二级碱基配对和/或碱基堆积结构实际上也不变。

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