STRUCTURE OF A NOVEL OXIDOREDUCTASE (TETX) IN TETRACYCLINE DRUG RESISTANCE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 自20世纪40年代发现以来，四环素类抗生素已被广泛用于治疗革兰氏阳性、革兰氏阴性和更不寻常的靶标，如衣原体和支原体。除了经典的药用用途外，金霉素和土霉素通常用于动物饲料中作为生长发育的亚治疗水平。随着四环素类抗生素成为生产范围最广的抗生素，耐药细菌的数量已经增加到几乎在临床和农业环境中无处不在的地步。耐药性通常是由药物外排(Teta)或核糖体保护(TetM和Teto)造成的。最近，人们发现了一种新的耐药酶机制--药物灭活(TetX)。我们已经找到了TetX的结晶条件，并正在申请收集MAD数据集的波束时间。在国际象棋F1中，非常小的天然晶体衍变到2.6安培(晶体是由同事拍摄的，但由于冻结不佳而无法收集数据)。自那以后，这些冻结问题已经得到解决。我们想要收集TetX蛋白的天然和MAD数据集以及药物复合体的浸泡。我们的研究特别重要，因为FDA批准的最新抗生素之一是泰格西尔或替吉环素，这是四环素类的一种，很容易被河豚毒素灭活。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Since their discovery in the 1940s, the tetracycline class of antibiotics has been widely used against a broad range of Gram positive, Gram negative and more unusual targets such as Chlamydia and mycoplasmas. In addition to classic medicinal applications, chlortetracycline and oxytetracycline are commonly used at sub-therapeutic levels in animal feed as growth. As the tetracyclines have risen to become the most widely produced antibiotic, the number of resistant bacteria has increased to the point that they are nearly ubiquitous in clinical and agricultural settings. Resistance is typically conferred by either efflux of the drug (tetA) or ribsomal protection (tetM and tetO). Recently, a new enzymatic mechanism of resistance was discovered through drug inactivation (tetX). We have found crystallization conditions for tetX and are requesting beamtime for the collection of a MAD data set. Very small native crystals diffracted to 2.6 A at CHESS F1 (the crystal was shot by a colleague but was unable to collect data because of poor freezes). These freezing issues have since been resolved. We would like to collect both a native and MAD data set of the tetX protein as well as a soak of the drug-complex. Our studies are particularly relevant as one of the most recent antibiotics approved by the FDA is Tygacil¿¿ or tigilcycline, a member of the tetracycline class is readily inactivated by tetX.