Toyama Hirohide, Inagaki Hideko, Matsushita Kazunobu, Anthony Christopher, Adachi Osao
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan.
Biochim Biophys Acta. 2003 Apr 11;1647(1-2):372-5. doi: 10.1016/s1570-9639(03)00097-9.
The largest of the gene clusters coding for proteins involved in methanol oxidation is the cluster mxaFJGIR(S)ACKLDEHB. Disruption of most of these genes leads to lack of growth on methanol. The previous results showed that the mutant lacking MxaD grows on methanol although at a low rate. This is explained by the low rate of methanol oxidation by whole cells. The specific activity of methanol dehydrogenase (MDH) is higher in the mutant but its electron acceptor (cytochrome c(L)) is unchanged. Using the purified proteins, it was shown that the rate of interaction of MDH and cytochrome c(L) was higher in the wild-type MDH containing some MxaD proteins, which was absent in the mutant MDH. It is suggested that the gene mxaD codes for the 17-kDa periplasmic protein that directly or indirectly stimulates the interaction between MDH and cytochrome c(L); its absence leads to a lower rate of respiration with methanol and therefore a lower growth rate on this substrate.
编码参与甲醇氧化的蛋白质的最大基因簇是mxaFJGIR(S)ACKLDEHB簇。这些基因中的大多数被破坏会导致在甲醇上无法生长。先前的结果表明,缺乏MxaD的突变体虽然生长速度较慢,但仍能在甲醇上生长。这可以通过全细胞甲醇氧化速率较低来解释。突变体中甲醇脱氢酶(MDH)的比活性较高,但其电子受体(细胞色素c(L))未发生变化。使用纯化的蛋白质表明,在含有一些MxaD蛋白的野生型MDH中,MDH与细胞色素c(L)的相互作用速率较高,而突变体MDH中则不存在这种情况。有人提出,基因mxaD编码17 kDa的周质蛋白,该蛋白直接或间接刺激MDH与细胞色素c(L)之间的相互作用;它的缺失导致甲醇呼吸速率降低,因此在该底物上的生长速率也降低。