Nakayama Y, Hayashi M, Unemoto T
Laboratory of Membrane Biochemistry, Faculty of Pharmaceutical Sciences, Chiba University, Japan.
FEBS Lett. 1998 Jan 30;422(2):240-2. doi: 10.1016/s0014-5793(98)00016-7.
We previously reported that the purified Na+-translocating NADH-quinone reductase (NQR) from the marine Vibrio alginolyticus is composed of three major subunits, alpha, beta and gamma. NQR operon was sequenced and was found to be composed of 6 structural genes. Among these genes, nqr1, nqr3 and nqr6 were identified to code for alpha-, gamma- and beta-subunits, respectively. The protein products from nqr2, nqr4 and nqr5, however, were not reported. The sequence data predicted that these three proteins are very hydrophobic and may be unusual in mobility and staining on SDS-PAGE. By modifying the detection method of proteins on SDS-PAGE, we could detect all six subunits encoded by NQR operon in the purified NQR complex. The open reading frame of each subunit was identified from its N-terminal amino acid sequence.
我们之前报道过,从海洋溶藻弧菌中纯化得到的钠转运型NADH-醌还原酶(NQR)由三个主要亚基α、β和γ组成。对NQR操纵子进行了测序,发现它由6个结构基因组成。在这些基因中,nqr1、nqr3和nqr6分别被鉴定为编码α-、γ-和β-亚基。然而,nqr2、nqr4和nqr5的蛋白质产物尚未见报道。序列数据预测这三种蛋白质具有很强的疏水性,在SDS-PAGE上的迁移率和染色情况可能不同寻常。通过改进SDS-PAGE上蛋白质的检测方法,我们能够在纯化的NQR复合物中检测到由NQR操纵子编码的所有六个亚基。从每个亚基的N端氨基酸序列鉴定出其开放阅读框。