OASIS to identify and analyze natural product biosynthesis

OASIS 识别和分析天然产物生物合成

基本信息

批准号：
8539383
负责人：
Michael D. Burkart
金额：
$ 27.71万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8539383
关键词：
Active Sites Address Adipocytes Affinity Labels Anabolism Antibiotics Area Attention Bacillus brevis Bacteria Bacterial Model Biological Biological Factors Case Study Collaborations Complex Development Dinophyceae Disease Enzymes Evaluation Research Food Genetic Genetic Techniques Genus Mycobacterium Goals Health Human Knowledge Laboratories Marines Medicine Metabolic Pathway Methods Mining Modeling Molecular Monitor Mycobacterium tuberculosis Mycobacterium ulcerans National Institute of Allergy and Infectious Disease Okadaic Acid Organism Pathway interactions Peat Peptides Pharmaceutical Preparations Pharmacologic Substance Production Proteins Proteome Proteomics Research Research Institute Science Shellfish Siderophores Solutions Source Sphagnum Moss Staging Stimulus Streptomyces System Systems Analysis Techniques Technology Toxin Tropical Disease Ulcer United States National Institutes of Health Virulence Factors activity-based protein profiling affinity labeling base design drug development interest killings marine organism mycobactins mycolactone non-ribosomal peptide synthase novel pathogen polyketide synthase programs protein aminoacid sequence public health relevance quorum sensing tool

项目摘要

DESCRIPTION (provided by applicant): Polyketides and non-ribosomal peptides are in a class of natural products important both as drug sources and as dangerous food toxins and virulence factors. While studies over the last two decades have provided substantial characterization of the modular synthases that produce these compounds at the genetic level, their understanding at the protein level is much less understood. Our laboratory has recently developed a proteomic platform, called an orthogonal active site identification system (OASIS), for identifying and quantifying natural product synthase enzymes. OASIS leverages activity based probes for enzyme enrichment in conjunction with highly sensitive peptide sequencing. To date, we have demonstrated the value of OASIS for the proteomic comparison of two model strains of Bacillus brevis. In this program, we apply this new tool to address topical problems in the discovery and study of modular synthases. Within this aim we have identified case studies that examine: antitubercular natural products from select sphagnum bog bacterial strains; the anticancer polyketide spirohexenolide B from Streptomyces platensis; the toxic virulence factor mycolactone from Mycobacterium ulcerans; and the shellfish toxin okadaic acid from the marine dinoflagellate Prorocentrum lima. Each of these studies offers solutions to important biomedical problems that have yet to be determined and remain currently unstudied due to the limitations of genetic techniques. The application of OASIS offers a new opportunity to evaluate these pathways through combining specific synthase-selective probes with proteomic technology, opening the tools of contemporary proteomics to natural product sciences. This program aims to bring the current state of natural product science into the proteomic era through the optimization and application of an orthogonal active site identification system (OASIS). The OASIS system provides new avenues into the study of natural product biosynthesis by enabling novel connectivity with mass spectral based proteomic analyses. Applications of the technique are examined within the context of natural product discovery, disease research, and the evaluation of dangerous toxins and virulence factors. PUBLIC HEALTH RELEVANCE: This program aims to bring the current state of natural product science into the proteomic era through the optimization and application of an orthogonal active site identification system (OASIS). The OASIS system provides new avenues into the study of natural product biosynthesis by enabling novel connectivity with mass spectral based proteomic analyses. Applications of the technique are examined within the context of natural product discovery, disease research, and the evaluation of dangerous toxins and virulence factors.

描述（由申请人提供）：聚酮化合物和非核糖体肽都在一类天然产物中，既重要，既重要，又是危险的食物毒素和毒力因子。尽管在过去的二十年中的研究提供了在遗传水平上产生这些化合物的模块化合酶的实质性表征，但它们在蛋白质水平上的理解知之甚少。我们的实验室最近开发了一个蛋白质组学平台，称为正交活性位点识别系统（OASIS），用于识别和量化天然产物合成酶。与高度敏感的肽测序结合使用绿洲基于活性的探针以富集酶富集。迄今为止，我们已经证明了绿洲对两种模型杆菌菌株的蛋白质组学比较的值。在此程序中，我们将此新工具应用于模块化合酶的发现和研究中的主题问题。在此目的中，我们确定了研究的案例研究：来自精选沼泽沼泽细菌菌株的抗结核天然产物；来自链霉菌链霉菌的抗癌聚酯螺旋甲甲烯醇B。溃疡分枝杆菌的毒性毒力因子菌根乳突；和贝类毒素冈达酸来自海洋鞭毛虫前lima。这些研究中的每一个都为重要的生物医学问题提供了解决方案，这些问题尚未确定，并且由于遗传技术的局限性，目前仍未研究。 OASIS的应用提供了一个新的机会，可以通过将特定的合酶选择性探针与蛋白质组学技术相结合，将当代蛋白质组学的工具与天然产品科学开放，以评估这些途径。该计划旨在通过优化和应用正交活动现场识别系统（OASIS）将自然产品科学的当前状态带入蛋白质组学时代。 OASIS系统通过通过基于质谱的蛋白质组学分析实现新的连通性来为自然产物生物合成的研究提供新的途径。在自然产品发现，疾病研究以及对危险毒素和毒力因素的评估的背景下检查了该技术的应用。公共卫生相关性：该计划旨在通过优化和应用正交主动现场识别系统（OASIS）将自然产品科学的当前状态带入蛋白质组学时代。 OASIS系统通过通过基于质谱的蛋白质组学分析实现新的连通性来为自然产物生物合成的研究提供新的途径。在自然产品发现，疾病研究以及对危险毒素和毒力因素的评估的背景下检查了该技术的应用。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Versatility of acyl-acyl carrier protein synthetases.

DOI：
10.1016/j.chembiol.2014.08.015
发表时间：
2014-10-23
期刊：
Chemistry & biology
影响因子：
0
作者：
Beld J;Finzel K;Burkart MD
通讯作者：
Burkart MD

The phosphopantetheinyl transferases: catalysis of a post-translational modification crucial for life.

DOI：
10.1039/c3np70054b
发表时间：
2014-01
期刊：
Natural product reports
影响因子：
11.9
作者：
Beld J;Sonnenschein EC;Vickery CR;Noel JP;Burkart MD
通讯作者：
Burkart MD

Explorations of catalytic domains in non-ribosomal peptide synthetase enzymology.

DOI：
10.1039/c2np20025b
发表时间：
2012-10
期刊：
Natural product reports
影响因子：
11.9
作者：
Hur GH;Vickery CR;Burkart MD
通讯作者：
Burkart MD

Visualizing the chain-flipping mechanism in fatty-acid biosynthesis.

DOI：
10.1002/anie.201408576
发表时间：
2014-12-22
期刊：
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
影响因子：
16.6
作者：
Beld, Joris;Cang, Hu;Burkart, Michael D.
通讯作者：
Burkart, Michael D.

Evolution of acyl-ACP-thioesterases and β-ketoacyl-ACP-synthases revealed by protein-protein interactions.

DOI：
10.1007/s10811-013-0203-4
发表时间：
2014-08-01
期刊：
JOURNAL OF APPLIED PHYCOLOGY
影响因子：
3.3
作者：
Beld, Joris;Blatti, Jillian L.;Behnke, Craig;Mendez, Michael;Burkart, Michael D.
通讯作者：
Burkart, Michael D.

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Michael D. Burkart其他文献

Data from mass spectrometry, NMR spectra, GC–MS of fatty acid esters produced by Lasiodiplodia theobromae

DOI：
10.1016/j.dib.2016.05.003
发表时间：
2016-09-01
期刊：
Data article
影响因子：
作者：
Carla C. Uranga;Joris Beld;Anthony Mrse;Iván Córdova-Guerrero;Michael D. Burkart;Rufina Hernández-Martínez
通讯作者：
Rufina Hernández-Martínez

The Complete Characterization of a Trapped ACYL Carrier Protein-Ketosynthase Complex

DOI：
10.1016/j.bpj.2019.11.1114
发表时间：
2020-02-07
期刊：
Conference abstract
影响因子：
作者：
Jeffrey T. Mindrebo;Laetitia E. Misson;Ashay Patel;Katia Charov;Joseph P. Noel;Michael D. Burkart
通讯作者：
Michael D. Burkart

Molecular Dynamics Simulations Reveal the Origins of the Distinct Activities and Substrate Selectivities of E. coli Dehydratases FabA and FabZ

DOI：
10.1016/j.bpj.2018.11.395
发表时间：
2019-02-15
期刊：
Conference abstract
影响因子：
作者：
Greg J. Dodge;Kara L. Jaremko;Ashay Patel;J. Andrew McCammon;Janet L. Smith;Michael D. Burkart
通讯作者：
Michael D. Burkart

Computational and Spectroscopic Investigation of Communication Mechanisms used by Acyl Carrier Proteins

DOI：
10.1016/j.bpj.2018.11.1035
发表时间：
2019-02-15
期刊：
Conference abstract
影响因子：
作者：
Terra Sztain-Pedone;Michael D. Burkart;James A. McCammon
通讯作者：
James A. McCammon

Elucidation of Protein-Protein Interactions Throughout E. coli Fatty Acid Biosynthesis

DOI：
10.1016/j.bpj.2019.11.1110
发表时间：
2020-02-07
期刊：
Conference abstract
影响因子：
作者：
Thomas G. Bartholow;Terra Sztain-Pedone;Ashay Patel;Ruben Abagyan;Michael D. Burkart
通讯作者：
Michael D. Burkart