Wall L, Rodriquez A, Meighen E
J Biol Chem. 1984 Feb 10;259(3):1409-14.
Acylation of extracts of Photobacterium phosphoreum at different stages of growth with [3H]tetradecanoic acid (+ATP) has shown that two polypeptides found in the fatty acid reductase complex, the fatty acid activating enzyme (50K) and the 34K polypeptide, were specifically labeled during induction of the luminescent system. An alternate method for in vitro acylation of polypeptides in the luminescent system was developed using tetradecanoyl-CoA. Both the 34K polypeptide and, to a lesser extent, the acyl-CoA reductase component (58K) in the complex, were acylated with [3H]tetradecanoyl-CoA. In contrast, the fatty acid activating enzyme (50K) was not labeled. Labeling of both the 34K and 58K polypeptides with [3H]tetradecanoyl-CoA as well as the acyl-CoA reductase activity in extracts paralleled the induction of luciferase during growth. Differential labeling of P. phosphoreum cells with [35S]methionine before luminescence induction and with [3H]methionine after the onset of luminescence followed by purification of luciferase and the polypeptides in the fatty acid reductase complex demonstrated that the alpha and beta subunits of luciferase and the 34K, 50K, and 58K polypeptides of the complex had 3H/35S ratios at least 7-fold higher than the constitutive proteins. These results give evidence that the synthesis of the component polypeptides of the fatty acid reductase are induced during the development of bioluminescence and may be under the same control as luciferase. The experiments also showed that P. phosphoreum may have the highest content of luciferase of any luminescent bacterium, constituting approximately 20% of the total soluble protein in extracts.
用[3H]十四烷酸（+ATP）对不同生长阶段的费氏弧菌提取物进行酰化，结果表明，在发光系统诱导过程中，脂肪酸还原酶复合物中的两种多肽，即脂肪酸活化酶（50K）和34K多肽，被特异性标记。利用十四烷酰辅酶A开发了一种在发光系统中对多肽进行体外酰化的替代方法。复合物中的34K多肽以及程度较轻的酰基辅酶A还原酶组分（58K）都被[3H]十四烷酰辅酶A酰化。相比之下，脂肪酸活化酶（50K）未被标记。用[3H]十四烷酰辅酶A对34K和58K多肽进行标记以及提取物中的酰基辅酶A还原酶活性，在生长过程中与荧光素酶的诱导情况平行。在发光诱导前用[35S]甲硫氨酸对费氏弧菌细胞进行差异标记，在发光开始后用[3H]甲硫氨酸进行标记，随后纯化荧光素酶和脂肪酸还原酶复合物中的多肽，结果表明荧光素酶的α和β亚基以及复合物中的34K、50K和58K多肽的3H/35S比值比组成型蛋白至少高7倍。这些结果证明，脂肪酸还原酶的组成多肽的合成在生物发光发育过程中被诱导，并且可能与荧光素酶受相同的调控。实验还表明，费氏弧菌可能是所有发光细菌中荧光素酶含量最高的，约占提取物中总可溶性蛋白的20%。
