Maïbèche-Coisne M, Boscameric M, Aragon S, Lafont R, Dauphin-Villemant C
Laboratoire d'Endocrinologie Moléculaire Comparée, Bât. A 5(ème) étage, 7 Quai St. Bernard, Paris, F-75005, France.
Biochem Biophys Res Commun. 2001 Mar 23;282(1):314-20. doi: 10.1006/bbrc.2001.4506.
Biochemical studies on ecdysteroid metabolism in arthropods suggest that aldoketoreductase enzymes (AKRs) may be involved in this pathway, but very few molecular data are available on these oxidoreductases in invertebrates. Looking for such enzymes in the crayfish Orconectes limosus, we have used a PCR strategy with primers deduced from a recent insect 3beta-reductase sequence, and from mammalian 5beta-reductase sequences. A full-length cDNA, corresponding to a putative AKR, was isolated from crayfish antennal gland. This cDNA contains an open-reading frame of 1008 bp, encoding a predicted protein of 336 amino acids. Northern blots indicated a restricted expression of the transcript in the antennal glands, quite constant during the molting cycle, and in situ hybridization demonstrated a strong expression of the transcript in the labyrinth. This is to date the first member of the AKRs superfamily characterized in a crustacean species, and the putative function of the corresponding enzyme is discussed.
对节肢动物蜕皮甾体代谢的生化研究表明，醛酮还原酶（AKRs）可能参与该途径，但关于这些氧化还原酶在无脊椎动物中的分子数据非常少。为了在小龙虾奥氏原螯虾中寻找此类酶，我们采用了一种PCR策略，使用从最近的昆虫3β-还原酶序列和哺乳动物5β-还原酶序列推导而来的引物。从小龙虾触角腺中分离出一个对应于假定AKR的全长cDNA。该cDNA包含一个1008 bp的开放阅读框，编码一个预测的336个氨基酸的蛋白质。Northern印迹表明该转录本在触角腺中表达受限，在蜕皮周期中相当稳定，原位杂交显示该转录本在迷路中强烈表达。这是迄今为止在甲壳类物种中鉴定出的AKRs超家族的首个成员，并对相应酶的假定功能进行了讨论。
