Litz Claudia, Helfmann Sarah, Gerhardt Stefan, Andrade Susana L A
Institut für Organische Chemie und Biochemie, Albert-Ludwigs-Universität Freiburg, Freiburg, Germany.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Feb 1;67(Pt 2):178-81. doi: 10.1107/S1744309110047482. Epub 2011 Jan 21.
GlnB and GlnK are ancient signalling proteins that play a crucial role in the regulation of nitrogen assimilation. Both protein types can be present in the same genome as either single or multiple copies. However, the gene product of glnK is always found in an operon together with an amt gene encoding an ammonium-transport (Amt) protein. Complex formation between GlnK and Amt blocks ammonium uptake and depends on the nitrogen level in the cell, which is regulated through the binding of specific effector molecules to GlnK. In particular, an ammonium shock to a cell culture previously starved in this nitrogen source or the binding of ATP to purified GlnK can stimulate effective complex formation. While the binding of ATP/ADP and 2-oxoglutarate (as a signal for low intracellular nitrogen) to GlnK have been reported and several GlnB/K protein structures are available, essential functional questions remain unanswered. Here, the crystal structure of A. fulgidus GlnK1 at 2.28 Å resolution and a comparison with the crystal structures of other GlnK proteins, in particular with that of its paralogue GlnK2 from the same organism, is reported.
谷氨酰胺结合蛋白B(GlnB)和谷氨酰胺结合蛋白K(GlnK)是古老的信号蛋白,在氮同化调节中发挥关键作用。这两种蛋白类型在同一基因组中可以以单拷贝或多拷贝形式存在。然而,glnK的基因产物总是与一个编码铵转运(Amt)蛋白的amt基因存在于同一个操纵子中。GlnK与Amt之间的复合物形成会阻止铵的吸收,并且这取决于细胞内的氮水平,而细胞内氮水平是通过特定效应分子与GlnK的结合来调节的。特别地,对先前在该氮源中饥饿的细胞培养物施加铵冲击,或者ATP与纯化的GlnK结合,都可以刺激有效的复合物形成。虽然已经报道了ATP/ADP和2-酮戊二酸(作为细胞内低氮信号)与GlnK的结合,并且有几种GlnB/K蛋白结构可供使用,但一些关键的功能问题仍未得到解答。本文报道了嗜热栖热菌GlnK1在2.28 Å分辨率下的晶体结构,并将其与其他GlnK蛋白的晶体结构进行了比较,特别是与其来自同一生物体的旁系同源物GlnK2的晶体结构进行了比较。