Suzuki Takayuki, Yoshimura Hidehisa, Hisabori Toru, Ohmori Masayuki
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro, Tokyo 153-8902, Japan.
FEBS Lett. 2004 Jul 30;571(1-3):154-60. doi: 10.1016/j.febslet.2004.06.074.
Two open reading frames (ORFs), alr0295 and alr2325, are found to encode putative cAMP receptor proteins (CRPs) in the genome of the filamentous cyanobacterium Anabaena sp. PCC 7120. These ORFs were named cAMP receptor protein-like gene A in Anabaena sp. PCC 7120 (ancrpA) and cAMP receptor protein-like gene B in Anabaena sp. PCC 7120 (ancrpB), respectively, and those translated products were investigated. The equilibrium dialysis measurements revealed that AnCrpA bound with cAMP specifically, while AnCrpB bound with both cAMP and cGMP, though the affinity for cGMP was weak. The binding affinity for cAMP of AnCrpA showed the lowest dissociation constant, approximately 0.8 microM, among bacterial CRPs. A gel mobility shift assay elucidated that AnCrpA and AnCrpB formed a complex with the consensus DNA sequence in the presence of cAMP, although AnCrpB did not have ordinary DNA-binding motifs.
在丝状蓝细菌鱼腥藻Anabaena sp. PCC 7120的基因组中,发现两个开放阅读框(ORF),即alr0295和alr2325,编码假定的环磷酸腺苷(cAMP)受体蛋白(CRP)。这些ORF分别被命名为鱼腥藻Anabaena sp. PCC 7120中的cAMP受体蛋白样基因A(ancrpA)和鱼腥藻Anabaena sp. PCC 7120中的cAMP受体蛋白样基因B(ancrpB),并对其翻译产物进行了研究。平衡透析测量结果显示,AnCrpA特异性结合cAMP,而AnCrpB则同时结合cAMP和环磷酸鸟苷(cGMP),尽管对cGMP的亲和力较弱。在细菌CRP中,AnCrpA对cAMP的结合亲和力表现出最低的解离常数,约为0.8微摩尔。凝胶迁移率变动分析表明,在cAMP存在的情况下,AnCrpA和AnCrpB与共有DNA序列形成复合物,尽管AnCrpB没有普通的DNA结合基序。