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使用更同源的培养基研究鸟氨酸氨甲酰基转移酶前体在体外向大鼠肝脏线粒体的转运。

In vitro transport of ornithine carbamoyltransferase precursor into rat liver mitochondria using a more homologous medium.

作者信息

Miralles V J, González-Bosch C, Marcote M J, Hernández-Yago J, Grisolía S

机构信息

Instituto de Investigaciones Citológicas de la Caja de Ahorros de Valencia, Centro Associado del CSIC, Spain.

出版信息

Arch Biochem Biophys. 1989 Nov 15;275(1):224-7. doi: 10.1016/0003-9861(89)90367-6.

Abstract

The precursor of ornithine carbamoyltransferase can be transported in vitro into rat liver mitochondria using the postmitochondrial supernatant from rat liver, a more homologous medium than the commonly used rabbit reticulocyte lysate. The transport of the precursor in the case of reticulocyte lysate requires a standard translation mixture. In the presence of the postmitochondrial supernatant the same is true. However, when the components of the translation mixture were added individually to the postmitochondrial supernatant, it was found that spermidine or spermine, at physiological concentrations, sufficed for the transport of the precursor of ornithine carbamoyltransferase. The activity of the postmitochondrial supernatant was inactivated by trypsin and slightly decreased by RNase treatment; it was not lost by dialysis or by heating at 100 degrees C.

摘要

鸟氨酸氨甲酰基转移酶的前体可利用大鼠肝脏的线粒体后上清液在体外转运至大鼠肝脏线粒体中,该上清液比常用的兔网织红细胞裂解液更具同源性。在前体转运方面,网织红细胞裂解液需要标准翻译混合物。在线粒体后上清液存在的情况下也是如此。然而,当将翻译混合物的成分单独添加到线粒体后上清液中时,发现生理浓度的亚精胺或精胺足以实现鸟氨酸氨甲酰基转移酶前体的转运。线粒体后上清液的活性可被胰蛋白酶灭活,经核糖核酸酶处理后略有降低;透析或在100℃加热不会使其活性丧失。

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