Sasso Severin, Okvist Mats, Roderer Kathrin, Gamper Marianne, Codoni Giosiana, Krengel Ute, Kast Peter
Laboratory of Organic Chemistry, ETH Zurich, CH-8093 Zurich, Switzerland.
EMBO J. 2009 Jul 22;28(14):2128-42. doi: 10.1038/emboj.2009.165. Epub 2009 Jun 25.
Chorismate mutase catalyzes a key step in the shikimate biosynthetic pathway towards phenylalanine and tyrosine. Curiously, the intracellular chorismate mutase of Mycobacterium tuberculosis (MtCM; Rv0948c) has poor activity and lacks prominent active-site residues. However, its catalytic efficiency increases >100-fold on addition of DAHP synthase (MtDS; Rv2178c), another shikimate-pathway enzyme. The 2.35 A crystal structure of the MtCM-MtDS complex bound to a transition-state analogue shows a central core formed by four MtDS subunits sandwiched between two MtCM dimers. Structural comparisons imply catalytic activation to be a consequence of the repositioning of MtCM active-site residues on binding to MtDS. The mutagenesis of the C-terminal extrusion of MtCM establishes conserved residues as part of the activation machinery. The chorismate-mutase activity of the complex, but not of MtCM alone, is inhibited synergistically by phenylalanine and tyrosine. The complex formation thus endows the shikimate pathway of M. tuberculosis with an important regulatory feature. Experimental evidence suggests that such non-covalent enzyme complexes comprising an AroQ(delta) subclass chorismate mutase like MtCM are abundant in the bacterial order Actinomycetales.
分支酸变位酶催化莽草酸生物合成途径中通向苯丙氨酸和酪氨酸的关键步骤。奇怪的是,结核分枝杆菌的细胞内分支酸变位酶(MtCM;Rv0948c)活性较差,且缺乏突出的活性位点残基。然而,添加另一种莽草酸途径酶——3-脱氧-D-阿拉伯庚酮糖酸-7-磷酸合酶(MtDS;Rv2178c)后,其催化效率提高了100倍以上。与过渡态类似物结合的MtCM-MtDS复合物的2.35 Å晶体结构显示,一个由四个MtDS亚基形成的中心核心夹在两个MtCM二聚体之间。结构比较表明,催化激活是MtCM活性位点残基在与MtDS结合时重新定位的结果。对MtCM C端突出部分的诱变确定了保守残基是激活机制的一部分。复合物的分支酸变位酶活性,而非单独的MtCM的活性,受到苯丙氨酸和酪氨酸的协同抑制。因此,复合物的形成赋予了结核分枝杆菌的莽草酸途径一个重要的调节特性。实验证据表明,在放线菌目中,像MtCM这样包含AroQ(δ)亚类分支酸变位酶的非共价酶复合物很丰富。