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腐胺对原代培养肝细胞中鸟氨酸脱羧酶合成及降解的影响。

Effects of putrescine on synthesis and degradation of ornithine decarboxylase in primary cultured hepatocytes.

作者信息

Kanamoto R, Utsunomiya K, Kameji T, Hayashi S

出版信息

Eur J Biochem. 1986 Feb 3;154(3):539-44. doi: 10.1111/j.1432-1033.1986.tb09432.x.

Abstract

Changes in both synthesis rate and degradation rate of ornithine decarboxylase (ODC) were pursued in primary cultures of adult rat hepatocytes during the process of ODC induction caused by asparagine and glucagon and also during the process of rapid ODC decay caused by putrescine. The synthesis rate of ODC was determined by [35S]methionine incorporation into the enzyme, which was separated afterwards by immunoprecipitation and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The degradation rate of ODC was determined by following the decay of prelabeled ODC. The enzyme induction caused by asparagine (10 mM) and glucagon (1 microM) was due both to an increase in the synthesis rate and to a decrease in the degradation rate. Addition of 10 mM putrescine caused a rapid decay of ODC activity, which was faster than ODC decay in the presence of cycloheximide. This rapid decay in ODC activity was accompanied by slightly slower decay in ODC protein, which was due both to partial suppression of ODC synthesis and to several fold acceleration of ODC degradation.

摘要

在成年大鼠肝细胞原代培养物中,研究了由天冬酰胺和胰高血糖素诱导鸟氨酸脱羧酶(ODC)过程中以及由腐胺导致ODC快速降解过程中ODC合成速率和降解速率的变化。ODC的合成速率通过将[35S]甲硫氨酸掺入该酶来确定,随后通过免疫沉淀和十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳进行分离。ODC的降解速率通过追踪预先标记的ODC的降解来确定。由天冬酰胺(10 mM)和胰高血糖素(1 μM)引起的酶诱导是由于合成速率增加和降解速率降低。添加10 mM腐胺导致ODC活性快速下降,这比在存在环己酰亚胺的情况下ODC的降解更快。ODC活性的这种快速下降伴随着ODC蛋白的稍慢降解,这是由于ODC合成的部分抑制和ODC降解的几倍加速所致。

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