Hou Wan-ru, Du Yu-jie, Chen Yu, Wu Xia, Peng Zheng-song, Yang Jun, Zhou Cai-Quan
College of Life Science, China West Normal University, Nanchong, Sichuan, China.
DNA Cell Biol. 2007 Nov;26(11):799-802. doi: 10.1089/dna.2007.0626.
Mitochondrial ATP synthase (F1Fo-ATPase) is regulated by an intrinsic ATPase inhibitor protein. In the present study, using RT-PCR combined with in silico cloning, we isolated and sequenced the cDNA encoding the inhibitor protein of the giant panda (Ailuropoda melanoleuca). The deduced protein sequence showed that the protein is composed of 106 amino acids and the estimated molecular weight of the ATPIF(1) protein is 12.32 kDa with an isoelectric point (pI) of 10.17. Alignment analysis revealed that the deduced protein sequence shares 66%, 78.3%, 66%, 72.6%, 77.4%, and 78.3% homology with that of Mus musculus, Pan troglodytes, Rattus norvegicus, Bos taurus, Macaca mulatta, and Homo sapiens, respectively. Topology prediction showed that there are three protein kinase C phosphorylation sites, one amidation site, three N-myristoylation sites, one casein kinase II phosphorylation site, and one tyrosine kinase phosphorylation site in the ATPase inhibitor. In particular, amino acids in the region between 39 and 72, which is the minimum sequence showing ATPase inhibitory activity, were highly conserved in the protein.
线粒体ATP合酶（F1Fo - ATP酶）受一种内在的ATP酶抑制蛋白调控。在本研究中，我们运用逆转录聚合酶链反应（RT-PCR）结合电子克隆技术，分离并测定了大熊猫（Ailuropoda melanoleuca）ATP酶抑制蛋白的编码cDNA序列。推导的蛋白质序列显示，该蛋白由106个氨基酸组成，ATPIF(1)蛋白的估计分子量为12.32 kDa，等电点（pI）为10.17。序列比对分析表明，推导的蛋白质序列与小家鼠、黑猩猩、褐家鼠、牛、猕猴和人类的序列同源性分别为66%、78.3%、66%、72.6%、77.4%和78.3%。拓扑结构预测显示，ATP酶抑制剂中有三个蛋白激酶C磷酸化位点、一个酰胺化位点、三个N-肉豆蔻酰化位点、一个酪蛋白激酶II磷酸化位点和一个酪氨酸激酶磷酸化位点。特别是，在39至72位之间的区域，即显示ATP酶抑制活性的最小序列中的氨基酸，在该蛋白中高度保守。
