Long Ashton Research Station, University of Bristol, BS18 9AF, Bristol, UK.
Planta. 1978 Jan;141(2):183-9. doi: 10.1007/BF00387887.
Induction of nitrate reductase (EC 1.6.6.1) activity was measured in Paul's Scarlet rose cell suspensions cultured in media containing nitrate (NO 3 (-) ) or urea (U) as nitrogen source, and with (+Mo) or without molybdenum (-Mo). There was a lag of 30 min during induction by NO 3 (-) in +Mo cultures but no lag occurred during induction after adding Mo to NO 3 (-) -Mo or to U-Mo cultures preincubated with NO 3 (-) . Actinomycin D, cycloheximide, and puromycin completely blocked induction by NO 3 (-) , but had no effect on the initial rate of induction by Mo. Cycloheximide and puromycin blocked induction by NO 3 (-) more quickly than actinomycin D. Induction by NO 3 (-) appeared to involve mRNA-dependent synthesis of apoprotein followed by rapid activation with molybdenum in intact cells independently of protein synthesis. Nitrate-induced apoprotein appeared less stable than the holoenzyme. When induced by NO 3 (-) in the absence of Mo, apoprotein concentration was about half the amount of maximally induced nitrate reductase. Cycloheximide stabilised preformed nitrate reductase which disappeared steadily in the presence of puromycin. Apoprotein was not stabilised by either antimetabolite.
诱导硝酸盐还原酶(EC 1.6.6.1)活性的测定方法是在含有硝酸盐(NO 3 (-) )或尿素(U)作为氮源的 Paul's Scarlet rose 细胞悬浮液中培养,并添加或不添加钼(Mo)。在+Mo 培养物中,NO 3 (-) 诱导硝酸盐还原酶活性有 30 分钟的滞后,但在向 NO 3 (-) -Mo 或预先用 NO 3 (-) 孵育的 U-Mo 培养物中添加 Mo 后,没有滞后发生。放线菌酮、环己酰亚胺和嘌呤霉素完全阻断了 NO 3 (-) 的诱导,但对 Mo 诱导的初始速率没有影响。环己酰亚胺和嘌呤霉素比放线菌酮更快地阻断了 NO 3 (-) 的诱导。NO 3 (-) 的诱导似乎涉及依赖于 mRNA 的脱辅基蛋白合成,然后在完整细胞中快速与钼激活,而不依赖于蛋白质合成。硝酸盐诱导的脱辅基蛋白似乎不如全酶稳定。当在没有 Mo 的情况下由 NO 3 (-) 诱导时,脱辅基蛋白浓度约为最大诱导的硝酸盐还原酶的一半。环己酰亚胺稳定了预先形成的硝酸盐还原酶,而在嘌呤霉素存在下,它会不断消失。两种代谢抑制剂都不能稳定脱辅基蛋白。