Cha Joo Young, Ishiwata Akihiro, Mobashery Shahriar
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, USA.
J Biol Chem. 2004 Apr 9;279(15):14917-21. doi: 10.1074/jbc.M400666200. Epub 2004 Jan 27.
Treponema pallidum, the causative agent of syphilis, is sensitive to penicillins. Yet, an abundant membrane-bound protein of this organism, Tp47, turns over penicillins. It is shown herein that the turnover process is a hydrolytic reaction that results in the corresponding penicilloates, products that have their beta-lactam bonds hydrolyzed. This is the reaction of beta-lactamases, bona fide resistance enzymes to beta-lactam antibiotics. Remarkably, the x-ray structure of Tp47 bears no resemblance to any other beta-lactamases or the related penicillin-binding proteins. Furthermore, evidence is presented that the reaction of Tp47 takes place in the absence of the zinc ion and does not involve intermediary acyl enzyme species. Hence, the beta-lactamase activity of Tp47 is the fifth known mechanism for turnover of beta-lactam antibiotics. Tp47 also exhibits a penicillin binding reaction, in the process of which the enzyme is covalently modified in the active site. The two reactions take place in two different active sites, and the events of the beta-lactamase activity are over 2,000-fold more rapid than the penicillin binding reaction. The level of beta-lactamase activity is high and is held back only by a strong product-inhibition component to the catalytic process. If natural selection would result in a mutant variant of Tp47 that overcomes product inhibition for the beta-lactamase activity, a novel bona fide resistance to penicillins will emerge in Treponema, which will be a disconcerting clinical development. The physiological functions of Tp47 are not known, but it is likely that this is at least a bifunctional enzyme involved in the processing of the Treponema peptidoglycan as a substrate.
梅毒的病原体梅毒螺旋体对青霉素敏感。然而,该生物体中一种丰富的膜结合蛋白Tp47会使青霉素失活。本文表明,失活过程是一种水解反应,会产生相应的青霉素酸盐,即β-内酰胺键被水解的产物。这是β-内酰胺酶的反应,β-内酰胺酶是对β-内酰胺抗生素真正有抗性的酶。值得注意的是,Tp47的X射线结构与任何其他β-内酰胺酶或相关的青霉素结合蛋白都不相似。此外,有证据表明Tp47的反应在没有锌离子的情况下发生,并且不涉及中间的酰基酶物种。因此,Tp47的β-内酰胺酶活性是已知的β-内酰胺抗生素失活的第五种机制。Tp47还表现出青霉素结合反应,在此过程中,酶在活性位点被共价修饰。这两种反应发生在两个不同的活性位点,β-内酰胺酶活性的反应速度比青霉素结合反应快2000倍以上。β-内酰胺酶活性水平很高,只是由于催化过程中强烈的产物抑制成分才受到抑制。如果自然选择导致Tp47的突变变体克服了β-内酰胺酶活性的产物抑制,梅毒螺旋体中将出现一种新的对青霉素的真正抗性,这将是一个令人不安的临床进展。Tp47的生理功能尚不清楚,但它可能至少是一种参与处理梅毒螺旋体肽聚糖作为底物的双功能酶。