Maeda Shin-ichi, Omata Tatsuo
Laboratory of Molecular Plant Physiology, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
J Bacteriol. 2009 May;191(10):3265-72. doi: 10.1128/JB.00013-09. Epub 2009 Mar 13.
In addition to the ATP-binding cassette (ABC)-type nitrate/nitrite-bispecific transporter, which has a high affinity for both substrates (K(m), approximately 1 microM), Synechococcus elongatus has an active nitrite transport system with an apparent K(m) (NO(2)(-)) value of 20 microM. We found that this activity depends on the cynABD genes, which encode a putative cyanate (NCO(-)) ABC-type transporter. Accordingly, nitrite transport by CynABD was competitively inhibited by NCO(-) with a K(i) value of 0.025 microM. The transporter was induced under conditions of nitrogen deficiency, and the induced cells showed a V(max) value of 11 to 13 micromol/mg of chlorophyll per h for cyanate or nitrite, which could supply approximately 30% of the amount of nitrogen required for optimum growth. Its relative specificity for the substrates and regulation at transcriptional and posttranslational levels suggested that the physiological role of the bispecific cyanate/nitrite transporter in S. elongatus is to allow nitrogen-deficient cells to assimilate low concentrations of cyanate in the medium. Its contribution to nitrite assimilation was significant in a mutant lacking the ABC-type nitrate/nitrite transporter, suggesting a possible role for CynABD in nitrite assimilation by cyanobacterial species that lack another high-affinity mechanism(s) for nitrite transport.
除了对两种底物(K(m),约1 microM)都具有高亲和力的ATP结合盒(ABC)型硝酸盐/亚硝酸盐双特异性转运蛋白外,聚球藻还具有一种活性亚硝酸盐转运系统,其表观K(m)(NO(2)(-))值为20 microM。我们发现这种活性依赖于cynABD基因,该基因编码一种假定的氰酸盐(NCO(-))ABC型转运蛋白。因此,CynABD介导的亚硝酸盐转运受到NCO(-)的竞争性抑制,K(i)值为0.025 microM。该转运蛋白在氮缺乏条件下被诱导,诱导后的细胞对氰酸盐或亚硝酸盐的V(max)值为每小时11至13微摩尔/毫克叶绿素,这可以提供最佳生长所需氮量的约30%。其对底物的相对特异性以及在转录和翻译后水平的调控表明,聚球藻中双特异性氰酸盐/亚硝酸盐转运蛋白的生理作用是使缺氮细胞能够同化培养基中的低浓度氰酸盐。在缺乏ABC型硝酸盐/亚硝酸盐转运蛋白的突变体中,其对亚硝酸盐同化的贡献显著,这表明CynABD在缺乏另一种高亲和力亚硝酸盐转运机制的蓝藻物种的亚硝酸盐同化中可能发挥作用。