Interrogation of Microbial Natural Product Methyltransferases

微生物天然产物甲基转移酶的研究

• 批准号: 10373164
• 负责人: Tony D. Davis
• 金额: $ 24.9万
• 依托单位: STATE UNIVERSITY OF NY,BINGHAMTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373164
• 关键词: Affinity; Anabolism; Anti-Bacterial Agents; Antibiotics; Binding; Biological; Carrier Proteins; Catalysis; Cell physiology; Cells; Clinic; Communication; Detection; Drug Design; Drug Targeting; Engineering; Enzymes; Epigenetic Process; Eukaryota; Fostering; Future; Gene Expression Regulation; Goals; In Vitro; Individual; Infection; Investigation; Life; Mediating; Methyltransferase; Molecular; Molecular Biology; Multiple Bacterial Drug Resistance; Mutagenesis; Natural Products; Nature; Neurons; Nucleic Acids; Organic Synthesis; Pathogenicity; Pathway interactions; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Play; Prevalence; Production; Property; Protein Engineering; Protein Methyltransferases; Proteins; Proteome; Research; Resistance; Role; S-Adenosylmethionine; Signal Transduction; Signaling Molecule; Specificity; Structure; Substrate Specificity; Virulence Factors; Work; activity-based protein profiling; alkyl group; analog; anti-cancer; base; biophysical techniques; cofactor; combinatorial; cost effective; design; in vivo; inhibitor/antagonist; insight; man; microbial; microorganism; novel; pathogen; pharmacophore; pressure; programs; protein complex; protein protein interaction; screening; small molecule; structural biology; therapeutic target; tool

PROJECT SUMMARY/ABSTRACT The increased prevalence of multi-drug resistant bacteria has led to an urgent unmet need to develop new an- timicrobial drugs. Targeting bacterial natural product biosynthesis has emerged as a promising avenue, due to the role of natural products as virulence factors, signaling factors, and agents of microbial warfare. However, to date, no drugs targeting enzymes in these biosynthetic pathways have reached the clinic. Hence, systematic investigations into key biosynthetic enzymes are necessary to understand fully their individual roles in virulence factor production. S-adenosyl-L-methionine (SAM)-dependent methyltransferases are ubiquitous throughout all domains of life and modify a diverse array of substrates, including proteins, nucleic acids, and natural products. In eukaryotes, methyltransferase activity modulates cellular processes such as epigenetic gene regulation and neuronal communication; consequently, these methyltransferases are extensively studied and have been validated as anticancer targets. In contrast, natural product methyltransferases have yet to be explored as therapeutic “tar- gets”, in spite of their important role in the biosynthesis of pathogenic virulence factors and pharmaceutically relevant compounds. During biosynthesis, natural product methyltransferases recognize substrates that are covalently tethered to a carrier protein; hence, successful methyl transfer is a function of both substrate recog- nition and proper protein-protein interactions between the methyltransferase and the carrier protein. The tran- sient nature of these protein-protein interactions makes targeting methyltransferases particularly challenging, yet elucidating the molecular basis of cofactor-protein, substrate-protein, and protein-protein recognition would offer a significant step towards advancing these efforts. Herein, our overall goal is to identify and engineer critical methyltransferase interactions during the biosynthe- sis of non-ribosomal peptide and polyketide-derived virulence factors by combining organic synthesis, ad- vanced molecular biology, and structural biology. We propose to (aim 1) develop small molecules to interro- gate cofactor and substrate methyltransferase activity, (aim 2) design tools to capture methyltransferase-carrier protein interactions, and (aim 3) develop selective activity-based probes for bacterial natural product methyl- transferases for detection and identification in the bacterial proteome. The insights from this work will be broad- ly significant by providing critical first steps in structure-based design of drugs targeting bacterial methyltrans- ferases, fostering combinatorial biosynthetic efforts of alkyl groups into unnatural products with precise regio-, stereo-, and chemoselectivity, and advancing medicinal chemistry efforts that alter biological and physico- chemical properties of privileged pharmacophores through chemoenzymatic synthesis.

项目摘要/摘要 多药耐药细菌的患病率提高导致迫切需要开发新的AN- 辛辣药物。靶向细菌天然产物生物合成已成为有前途的途径 天然产物作为病毒因素，信号因子和微生物战的药物的作用。但是，要 日期，没有针对这些生物合成途径中酶的药物已经到达诊所。因此，系统 对关键生物合成酶的研究是必要的 因子产生。 S-腺苷-l-甲硫代（SAM）依赖性甲基转移酶在生命的所有领域中无处不在 并修改潜水员的底物，包括蛋白质，核酸和天然产物。在真核生物中， 甲基转移酶活性调节细胞过程，例如表观遗传基因调节和神经元 沟通;因此，这些甲基转移酶是广泛研究的，已被证实为 抗癌目标。相比之下，天然产物甲基转移酶尚待探索，因为 “得到”，尽管它们在致病病毒因子和药物的生物合成中起着重要作用 相关化合物。在生物合成期间，天然产物甲基转移酶识别底物 共价绑在载体蛋白上；因此，成功的甲基转移是两个底物的函数 甲基转移酶与载体蛋白之间的核定和适当的蛋白质蛋白相互作用。 tran- 这些蛋白质蛋白相互作用的良性性质使靶向甲基转移酶特别具有挑战性， 然而，阐明辅因子 - 蛋白质，底物 - 蛋​​白质和蛋白质蛋白识别的分子基础将 朝着推进这些努力迈出了重要一步。 在此，我们的总体目标是在生物合成期间识别和设计关键的甲基转移酶相互作用 - 通过结合有机合成，AD- 分子生物学和结构生物学。我们建议（AIM 1）开发小分子到中间 栅极辅因子和底物甲基转移酶活性（AIM 2）设计工具以捕获甲基转移酶载体 蛋白质相互作用，（AIM 3）为细菌天然产物甲基 - 基于选择性活性的问题 - 在细菌蛋白质组中检测和鉴定的转移酶。这项工作的见解将是广泛的 - 通过在基于结构的基于结构的靶向甲基转移的药物设计中提供关键的第一步，这很重要 铁酶，将烷基组合的联合生物合成努力促进具有精确区域的非天然产物 立体和化学选择性以及进步的医学化学努力，以改变生物学和物理学 通过化学酶合成的特权药理化学特性。