Peptide Deformylase: Mechanism and Inhibitor Design

项目描述（由申请人提供）：本项目的目标是（1） 为了表征使N-甲酰化肽（肽）去甲酰化的酶 脱甲酰基酶（PDF）、甲酰甲硫氨酸氨基肽酶（fMAP）和甲酰甲硫氨酸 脱甲酰基酶（fetoxylase，fetoxylase）），和（2）评估PDF作为抗微生物药物的新靶点 药物设计在细菌和真核细胞器中， 与N-甲酰甲硫氨酸因此，所有新合成的多肽在 细菌和细胞器携带N-末端甲酰基。以下 在翻译起始时，PDF从绝大多数中除去N-甲酰基基团， 细菌蛋白质。作为细菌的一种基本活性，PDF正在被 作为抗菌药物设计的新目标。最近，Genomic 测序已经揭示了某些真核生物包括人中的PDF样序列， 提出了一些关于PDF作为药物靶点的有效性的问题。一个相关 问题是PDF抑制剂的治疗是否会导致累积 的N-甲酰肽和炎症的患者，因为一些N-甲酰肽， 肽（例如，f-MLF）是有效的趋化剂。为了解决这些问题， 该项目提出了五个具体目标。具体目标1是执行 定量分析了E.线圈PDF。 活性位点中的保守残基将被突变，并且动力学、光谱、 并进行结构表征。具体目标2是克隆 并表征来自人和恶性疟原虫的PDF样序列， 疟疾的病原体。目标是确定这些 序列实际上编码功能性PDF及其生理功能。 具体目标3是进一步提高PDF的效力和特异性 针对细菌PDF的抑制剂，设计针对P. 恶性疟原虫PDF，并评估PDF作为抗疟药物的潜在靶点 药物设计具体目标4是设计可以选择性地 由PDF激活。预期这样的前药更特异（因此更少）。 有毒）并且细菌难以产生耐药性。具体目标5是 从大鼠中纯化、克隆和表征fMAP和fMAP。这些酶 被认为是负责灭活趋化肽释放的 共生细菌并预防哺乳动物的炎症反应。

DESCRIPTION (provided by the applicant): The objective of this project is (1) to characterize the enzymes which deformylate N-formylated peptides (peptide deformylase (PDF), formylmethionine aminopeptidase (fMAP), and formylmethionine deformylase (fMDF)), and (2) to assess PDF as a novel target for antimicrobial drug design. In bacteria and eukaryotic organelles, protein synthesis initiates with N-formylmethionine. Consequently, all newly synthesized polypeptides in bacteria and organelles carry an N-terminal formyl group. Following translational initiation, PDF removes the N-formyl group from the vast majority of bacterial proteins. As an essential activity in bacteria, PDF is being pursued as a new target for antibacterial drug design. Recently, genomic sequencing has revealed PDF-like sequences in certain eukaryotes including man, raising some questions about the validity of PDF as a drug target. A related issue is whether treatment with PDF inhibitors would result in the accumulation of N-formyl peptides and inflammation in a patient, since some of the N-formyl peptides (e.g., f-MLF) are potent chemotactic agents. To address these issues, five specific aims are proposed in this project. Specific Aim 1 is to perform quantitative analyses of the structure-function relationship of E. coil PDF. Conserved residues in the active site will be mutated and kinetic, spectral, and structural characterization will be conducted. Specific Aim 2 is to clone and characterize the PDF-like sequences from human and Plasmodium falciparum, the causative agent of malaria. The goal is to determine whether these sequences actually code for functional PDF's and their physiological functions. Specific Aim 3 is to further improve the potency and specificity of PDF inhibitors against bacterial PDF, to design potent inhibitors against P. falciparum PDF, and to evaluate PDF as a potential target for antimalarial drug design. Specific Aim 4 is to design prodrugs that can be selectively activated by PDF. Such prodrugs are expected to be more specific (thus less toxic) and difficult for bacteria to develop resistance. Specific Aim 5 is to purify, clone, and characterize fMAP and fMDF from rats. These enzymes are thought to be responsible for inactivating the chemotactic peptides released by commensal bacteria and preventing inflammatory responses in mammals.