The Promiscuity of EpoC Towards Polymer Bound Substrates

EpoC 对聚合物结合基底的混杂性

• 批准号: 6984779
• 负责人: Zachary David Aron
• 金额: $ 4.6万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-03 至 2007-09-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6984779
• 关键词: acid aminoacid ligase; biological products; coenzyme A; combinatorial chemistry; enzyme activity; enzyme biosynthesis; enzyme mechanism; enzyme model; enzyme substrate; epothilon; hydroxyl group; macrolide antibiotics; mass spectrometry; peptide chemical synthesis; peptide hormone analog; peptide library; polyketide synthase; postdoctoral investigator; protein protein interaction; protein structure function; stereochemistry; thioester

DESCRIPTION (provided by applicant): Small molecule peptide, polyketide and hybrid polyketide/polypeptide natural products are characterized by their potent biological activities and modular biosyntheses. These compounds typically are generated by modular polyketide synthase (PKS), nonribosomal peptide synthetase (NRPS) and hybrid NRPS/PKS enzymes. To date, there has been limited study of the NRPS/PKS interface of hybrid NRPS/PKS enzymes. This proposal seeks to examine the NRPS/PKS interface of epothilone synthase by exploring the promiscuity of the ketosynthase (KS) domain of epoC towards solid phase bound peptide substrates. EpoC, which sits at the hybrid NRPS/PKS interface of epothilone synthase, was chosen for this study because of the pharmaceutical significance of the epothilones and the extensive body of work regarding their biosynthesis. By applying this technology to libraries of peptide substrates a detailed understanding of the substrate specificity of epoC will be developed, opening the door to reengineering epothilone biosynthesis to generation of novel epothilone derivatives. Also, by fusing epoC to additional PKS modules, combinatorial libraries of hybrid polypeptide/polyketide molecules can be accessed for drug discovery by semibiosynthesis.

描述（由申请人提供）： 小分子肽、聚酮化合物和杂化聚酮化合物/多肽天然产物的特征在于其有效的生物活性和模块化生物合成。这些化合物通常由模块化聚酮合酶 (PKS)、非核糖体肽合成酶 (NRPS) 和杂合 NRPS/PKS 酶产生。迄今为止，对杂合 NRPS/PKS 酶的 NRPS/PKS 界面的研究还很有限。该提案旨在通过探索 epoC 的酮合酶 (KS) 结构域与固相结合肽底物的混杂性来检查埃坡霉素合酶的 NRPS/PKS 界面。 EpoC 位于埃博霉素合酶的杂合 NRPS/PKS 界面，由于埃博霉素的药学意义以及与其生物合成相关的大量工作，被选用于本研究。通过将该技术应用于肽底物库，将详细了解 epoC 的底物特异性，为重新设计埃博霉素生物合成以生成新型埃博霉素衍生物打开了大门。此外，通过将 epoC 与其他 PKS 模块融合，可以访问杂合多肽/聚酮化合物分子的组合文库，用于通过半生物合成进行药物发现。