Bognar A, Michaliszyn G, Meighen E
Can J Biochem. 1978 Jun;56(6):605-10. doi: 10.1139/o78-091.
The proteins of the bioluminescent bacterium Beneckea harveyi have been labelled with [3H]leucine prior to the induction of bioluminescence, and with [14C]leucine during the development of the bioluminescent system. An aliphatic aldehyde dehydrogenase and a NAD(P)H:flavin oxidoreductase, two enzymes that may be directly involved in the metabolism of the substrates (aldehyde, FMNH2) for the luminescent reaction catalyzed by luciferase, were purified and the isotope ratios of their respective polypeptide chains determined after sodium dodecyl sufate gel electrophoresis. A comparison of these isotope ratios to (a) the isotope ratios of the induced polypeptide chains of luciferase, purified in the same experiment, and (b) the average isotope ratio for the proteins synthesized in concert with growth has provided direct evidence that the synthesis of aldehyde dehydrogenase but not NAD(P)H:flavin oxidoreductase is induced during the development of bioluminescence.
在诱导生物发光之前，用[³H]亮氨酸标记哈维氏贝内克氏菌的蛋白质；在生物发光系统发育过程中，用[¹⁴C]亮氨酸进行标记。纯化了脂肪族醛脱氢酶和NAD(P)H：黄素氧化还原酶这两种可能直接参与底物（醛、FMNH₂）代谢的酶，这些底物参与由荧光素酶催化的发光反应，在十二烷基硫酸钠凝胶电泳后，测定了它们各自多肽链的同位素比率。将这些同位素比率与（a）在同一实验中纯化的荧光素酶诱导多肽链的同位素比率，以及（b）与生长同时合成的蛋白质的平均同位素比率进行比较，提供了直接证据，表明在生物发光发育过程中，醛脱氢酶的合成被诱导，而NAD(P)H：黄素氧化还原酶的合成未被诱导。
