Shi Xiaobing, Rao Narayana N, Kornberg Arthur
Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305-5307, USA.
Proc Natl Acad Sci U S A. 2004 Dec 7;101(49):17061-5. doi: 10.1073/pnas.0407787101. Epub 2004 Nov 30.
Chains of inorganic polyphosphate (poly-P) with hundreds of P(i) residues linked by phosphoanhydride bonds, as in ATP, are found in every bacterial, fungal, plant, and animal cell, in which they perform various functions. In the spore-forming Bacillus cereus, we have identified three principal enzymes and genes involved in the metabolism of poly-P, namely, (i) poly-P kinase (PPK), which synthesizes poly-P reversibly from ATP, (ii) exopolyphosphatase (PPX), which hydrolyzes poly-P to P(i), and (iii) poly-P/AMP phosphotransferase (PAP), which uses poly-P as a donor to convert AMP to ADP, reversibly. In the null mutant of ppk, poly-P levels are reduced to <5% of the WT; in the ppx mutant, the PPK activity is elevated 10-fold, and the accumulation of poly-P is elevated approximately 1,000-fold. All of the null mutants of ppk, ppx, and pap showed defects in motility and biofilm formation, but sporulation efficiency was impaired only in the ppx mutant. These enzymes and genes in B. cereus are nearly identical to those in the very closely related pathogen Bacillus anthracis, and, thus, they may provide attractive targets for the treatment of anthrax.
无机多聚磷酸盐(多聚-P)链含有数百个通过磷酸酐键相连的无机磷酸(P(i))残基,如同ATP中的情况一样,存在于每一个细菌、真菌、植物和动物细胞中,并在其中发挥各种功能。在形成芽孢的蜡样芽孢杆菌中,我们已经鉴定出三种参与多聚-P代谢的主要酶和基因,即:(i)多聚-P激酶(PPK),它能从ATP可逆地合成多聚-P;(ii)外多聚磷酸酶(PPX),它将多聚-P水解为P(i);以及(iii)多聚-P/AMP磷酸转移酶(PAP),它以多聚-P作为供体将AMP可逆地转化为ADP。在ppk基因的缺失突变体中,多聚-P水平降至野生型的<5%;在ppx突变体中,PPK活性升高10倍,多聚-P的积累升高约1000倍。ppk、ppx和pap的所有缺失突变体在运动性和生物膜形成方面均表现出缺陷,但只有ppx突变体的芽孢形成效率受损。蜡样芽孢杆菌中的这些酶和基因与密切相关的病原体炭疽芽孢杆菌中的那些酶和基因几乎相同,因此,它们可能为炭疽病的治疗提供有吸引力的靶点。