Crystallographic Studies of Polyphosphate Kinase

超出所提供的空间。 聚磷酸酯（poly（P））是由数百个正磷酸（Pi）残基通过高能磷酸酐键连接的线性聚合物。许多证据表明，聚（P）在应激和营养缺乏的调节反应中起着关键作用。聚磷酸激酶（PPK）是一种负责从ATP可逆合成聚（P）的酶。最近已经表明，PPK对于包括铜绿假单胞菌（P.aeruginosa）在内的许多病原微生物的生物膜发育、群体感应、运动性和毒力因子的释放是必需的。PPK基因的敲除导致病原微生物活力的丧失。因此，PPK特异性抑制剂可能成为一种新的抗生素家族，这对于克服囊性纤维化和其他免疫缺陷患者的抗生素耐药性非常有用，并且可以在打击生物恐怖主义中发挥作用。 PPK是一种膜相关酶，与其他蛋白质没有明显的序列同源性。PPK系列没有三维结构信息。我们最近纯化并结晶了全长E。coliPPK的晶体结构，并获得了晶体结构测定所需的初始相。该建议的主要目标是确定PPK及其与各种底物和反应中间体模拟物的复合物的晶体结构。我们的结构研究将有助于确定PPK催化机制，并为药物设计提供见解。城市、州）性能现场=

EXCEED THE SPACE PROVIDED. Polyphosphate (poly(P)) is a linear polymer of hundreds of orthophosphate (Pi) residues linked by high-energy phosphoanhydride bonds. Many lines of evidence indicate that poly(P) plays a critical role in regulatory responses to stresses and nutritional deficiencies. Polyphosphate kinase (PPK) is an enzyme responsible for the reversible synthesis of poly(P) from ATP. It has been shown recently that PPK is essential for biofilm development, quorum sensing, motility and release of virulence factors of many pathogenic microorganisms, including P. aeruginosa. Knockout of the PPK gene leads to the loss of viability of the pathogenic microorganism. Thus PPK specific inhibitors could become a novel family of antibiotics, which can be very useful to overcome antibiotic resistance in cystic fibrosis and other immunodeficient patients, and could play a role in combating bioterrorism. PPK is a membrane-associated enzyme that does not have apparent sequence homology with other proteins. No three-dimensional structural information is available for the PPK family. We have recently purified and crystallized the full-length E. coli PPK, and the initial phases for structure determination by X- ray crystallography have been obtained. The main goal of this proposal is the determination of crystal structures of PPK and its complexes with various substrates and reaction intermediate mimics. Our structural studies will aid in determining the PPK catalytic mechanism and provide insights into drug design. city, state) PERFORMANCE SITE ========================================Section End===========================================