Satoh Motohiko, Koyama Noriyuki
Faculty of Science, Department of Chemistry, Chiba University, Yayoi, Inage-ku, Chiba 263-8522, Japan.
Anaerobe. 2005 Feb-Apr;11(1-2):115-21. doi: 10.1016/j.anaerobe.2004.08.003.
The structural genes for A and B subunits of the V-type Na(+)-ATPase from a facultatively anaerobic alkaliphile (Amphibacillus sp.), strain M-12, were cloned and sequenced. Transformation of Escherichia coli with the genes overexpressed two proteins, which crossreacted with an antiserum against A and B subunits of the V-type Na(+)-ATPase from Enterococcus hirae. The deduced amino acid sequence (594 amino acids; Mr, 66,144) of A subunit of the M-12 enzyme exhibited 73%, 51%, 49% and 53% identities with those of V-type ATPases from E. hirae, Thermus thermophilus, Neurospora crassa and Drosophila melanogaster, respectively. The amino acid sequence (458 amino acids; Mr, 51,308) of B subunit of the M-12 enzyme was 74%, 53%, 52% and 54% identical with those of the ATPases from E. hirae, T. thermophilus, N. crassa and D. melanogaster, respectively. The fact indicates that the amino acid sequences of A and B subunits of the M-12 enzyme exhibit significantly higher homologies with those of the E. hirae Na(+)-ATPase as compared with those of the H(+)-ATPases from T. thermophilus, N. crassa and D. melanogaster.
对兼性厌氧嗜碱菌（两栖芽孢杆菌属）M-12菌株V型Na(+)-ATP酶A和B亚基的结构基因进行了克隆和测序。用这些基因转化大肠杆菌后，两种蛋白质得到过量表达，它们能与针对平肠球菌V型Na(+)-ATP酶A和B亚基的抗血清发生交叉反应。M-12酶A亚基推导的氨基酸序列（594个氨基酸；Mr，66,144）与平肠球菌、嗜热栖热菌、粗糙脉孢菌和黑腹果蝇V型ATP酶的氨基酸序列分别具有73%、51%、49%和53%的同一性。M-12酶B亚基的氨基酸序列（458个氨基酸；Mr，51,308）与平肠球菌、嗜热栖热菌、粗糙脉孢菌和黑腹果蝇ATP酶的氨基酸序列分别具有74%、53%、52%和54%的同一性。这一事实表明，与嗜热栖热菌、粗糙脉孢菌和黑腹果蝇的H(+)-ATP酶相比，M-12酶A和B亚基的氨基酸序列与平肠球菌Na(+)-ATP酶的氨基酸序列具有更高的同源性。
