The Arabidopsis thaliana K+-uptake permease 7 (AtKUP7) contains a functional cytosolic adenylate cyclase catalytic centre
The Arabidopsis thaliana K+-uptake permease 7 (AtKUP7) contains a functional cytosolic adenylate cyclase catalytic centre
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DOI：
10.1016/j.febslet.2015.11.038
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发表时间：
2015-12-21
期刊：
影响因子：
3.5
通讯作者：
Gehring, Chris
中科院分区：
文献类型：
--
作者：
Al-Younis, Inas;Wong, Aloysius;Gehring, Chris
腺苷酸环化酶（AC）催化从腺苷5 '-三磷酸（ATP）形成第二信使环腺苷3'，5 '-单磷酸（cAMP）。虽然cAMP越来越多地被认为是高等植物中重要的信号分子，但AC仍然有些难以捉摸。在这里，我们使用了一个搜索基序来自实验测试鸟苷酸环化酶（GC），取代的底物特异性所必需的残基，并确定了拟南芥K+-摄取通透酶7（AtKUP 7）作为几个候选AC之一。首先，我们表明，重组N-末端，AtKUP 7（1-100）的胞质结构域能够补充大肠杆菌中的AC缺陷型突变体cyaA，从而恢复乳糖的发酵，其次，我们用酶免疫测定和质谱证明，重组AtKUP 7（1-100）在体外产生cAMP。(C)2015年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
Adenylate cyclases (ACs) catalyse the formation of the second messenger cyclic adenosine 3',5'-monophosphate (cAMP) from adenosine 5'-triphosphate (ATP). Although cAMP is increasingly recognised as an important signalling molecule in higher plants, ACs have remained somewhat elusive. Here we used a search motif derived from experimentally tested guanylyl cyclases (GCs), substituted the residues essential for substrate specificity and identified the Arabidopsis thaliana K+-uptake permease 7 (AtKUP7) as one of several candidate ACs. Firstly, we show that a recombinant N-terminal, cytosolic domain of AtKUP7(1-100) is able to complement the AC-deficient mutant cyaA in Escherichia coli and thus restoring the fermentation of lactose, and secondly, we demonstrate with both enzyme immunoassays and mass spectrometry that a recombinant AtKUP7(1-100) generates cAMP in vitro. (C) 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.