Sherman T D, Funkhouser E A
Department of Biochemistry and Biophysics, Texas A&M University, College Station 77843.
Arch Biochem Biophys. 1989 Nov 1;274(2):525-31. doi: 10.1016/0003-9861(89)90466-9.
Assimilatory nitrate reductase is an inducible, eukaryotic enzyme that responds to a variety of environmental cues. When higher plants and green algae are grown with ammonia as a nitrogen source, low levels of nitrate reductase activity are present. Transfer to nitrate-containing medium is accompanied by substantial increase of nitrate reductase activity. Here it is shown immunologically that, in the green algae Chlorella vulgaris, nitrate reductase protein is over-produced as activity appears during induction. Immunoreactive protein is also found in cells grown on ammonia. Low levels of translatable mRNA for nitrate reductase are present in ammonia-grown cells. These data suggest that: (i) nitrate reductase appearance is controlled primarily on a transcriptional level, but that transcription is not completely halted under repressing conditions; (ii) there is an overproduction of nitrate reductase protein early during the induction period as previously suggested; and (iii) nascent protein, from in vitro translation, is of approximately the same molecular size as the nitrate reductase subunit and therefore little posttranslational modification is necessary to generate the functional enzyme. Insertion of cofactors and assembly are probably the only post-translational events.
同化型硝酸还原酶是一种可诱导的真核酶,它对多种环境信号作出反应。当高等植物和绿藻以氨作为氮源生长时,硝酸还原酶活性水平较低。转移到含硝酸盐的培养基中会伴随着硝酸还原酶活性的大幅增加。在此通过免疫方法表明,在绿藻小球藻中,随着诱导过程中活性的出现,硝酸还原酶蛋白过量产生。在以氨为氮源生长的细胞中也发现了免疫反应性蛋白。在以氨为氮源生长的细胞中存在低水平的可翻译的硝酸还原酶mRNA。这些数据表明:(i)硝酸还原酶的出现主要在转录水平上受到控制,但在抑制条件下转录并未完全停止;(ii)如先前所述,在诱导期早期硝酸还原酶蛋白过量产生;(iii)体外翻译产生的新生蛋白与硝酸还原酶亚基的分子大小大致相同,因此产生功能酶几乎不需要翻译后修饰。辅因子的插入和组装可能是唯一的翻译后事件。