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豌豆根多核糖体与细胞骨架的共沉降

Cosedimentation of pea root polysomes with the cytoskeleton.

作者信息

You W, Abe S, Davies E

机构信息

Molecular Probes, Eugene, OR.

出版信息

Cell Biol Int Rep. 1992 Jul;16(7):663-73. doi: 10.1016/s0309-1651(06)80008-1.

Abstract

When conventional, high ionic strength buffers were used for the isolation of polysomes from pea roots, only about 10% were retained in the detergent-insoluble pellet, and they were not degraded by endogenous RNase. A low ionic strength, cytoskeleton-stabilizing buffer increased retention to 60%, but polysomes were severely degraded. The RNase inhibitors, ribonucleoside-vanadyl complexes, heparin, KCl and ammonium sulphate lessened degradation but caused release, while Tris-HCl at 15-25 mM was able to prevent degradation without causing release. Cosedimentation of polysomes with the cytoskeleton is not an artefact of adsorption or trapping since isolated polysomes labelled through their nascent polypeptides and added to unfractionated tissue were not retained in the cytoskeletal pellet.

摘要

当使用常规的高离子强度缓冲液从豌豆根中分离多核糖体时,只有约10%保留在去污剂不溶性沉淀中,并且它们不会被内源性核糖核酸酶降解。低离子强度、稳定细胞骨架的缓冲液可将保留率提高到60%,但多核糖体会严重降解。核糖核酸酶抑制剂、核糖核苷-钒复合物、肝素、氯化钾和硫酸铵可减少降解,但会导致释放,而15-25 mM的Tris-HCl能够防止降解而不引起释放。多核糖体与细胞骨架的共沉降不是吸附或捕获的假象,因为通过其新生多肽标记并添加到未分级组织中的分离多核糖体不会保留在细胞骨架沉淀中。

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