Suzuki Yuusuke, Ueno Sumie, Ohnuma Rieko, Koyama Noriyuki
Department of Chemistry, Faculty of Science, Chiba University, Yayoi, Inage-ku, Chiba 263-8522, Japan.
Biochim Biophys Acta. 2005 Mar 10;1727(3):162-8. doi: 10.1016/j.bbaexp.2004.12.008. Epub 2005 Jan 12.
Cloning and sequencing of the gene encoding a P-type Na(+)-ATPase of a facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum, were conducted. The structural gene was composed of 2628 nucleotides. The deduced amino acid sequence (876 amino acid residues; Mr, 96,664) suggested that the enzyme possesses 10 membrane-spanning regions. When the amino acid sequences of the four putative membrane regions, M4, M5, M6 and M8, of BL77/1 ATPase were aligned with those of fungal Na(+)-ATPase, Na(+)/K(+)-ATPase, H(+)-ATPases and sarcoplasmic reticulum Ca(2+)-ATPase, it exhibited the highest homology with Ca(2+)-ATPase except M5 region. By the transformation of Escherichia coli with the expression vector (pQE30) containing the ATPase gene, the enzyme was functionally expressed in E. coli membranes.
对兼性厌氧嗜碱菌橙色栖热菌（Exiguobacterium aurantiacum）编码P型Na(+) -ATP酶的基因进行了克隆和测序。该结构基因由2628个核苷酸组成。推导的氨基酸序列（876个氨基酸残基；Mr，96,664）表明该酶具有10个跨膜区域。当将BL77/1 ATP酶的四个推定膜区域M4、M5、M6和M8的氨基酸序列与真菌Na(+) -ATP酶、Na(+)/K(+) -ATP酶、H(+) -ATP酶和肌浆网Ca(2+) -ATP酶的氨基酸序列进行比对时，除M5区域外，它与Ca(2+) -ATP酶具有最高的同源性。通过用含有ATP酶基因的表达载体（pQE30）转化大肠杆菌，该酶在大肠杆菌膜中实现了功能性表达。
