来自皮克罗米辛多基酸合成酶的脱水酶域的功能性表征
Yang Li1, Greg J Dodge2, William D Fiers1
1†Department of Medicinal Chemistry, College of Pharmacy, University of Minnesota, Minneapolis, Minnesota 55455, United States.
Journal of the American Chemical Society
|June 2, 2015
概括
这项研究特征PikDH2脱水酶 (DH) 域从多基化合成酶 (PKS) 中,揭示了它创造不和化合物的机制. 这项工作可以通过代谢工程发现新的天然产品.
科学领域:
- 生物化学 生物化学
- 代谢工程是代谢工程.
- 自然产品 发现 发现 发现
背景情况:
- 聚基酸合成酶 (PKS) 途径对于天然产品的生物合成至关重要.
- 由于测定局限性,PKS中的脱水酶 (DH) 域的理解较少.
- PKS路径的代谢工程为新型天然产品的发现提供了一个有前途的途径.
研究的目的:
- 首次对PKS DH域 (PikDH2) 进行稳定状态运动分析.
- 阐明PikDH2.2的基质特异性和催化机制.
- 为未来对PKS DH领域的结构研究奠定基础.
主要方法:
- 使用液体染色学-双重质谱法 (LC-MS/MS) 进行产品定量.
- 采用了一系列合成的三基基底与非水解的乙烯链接.
- 进行了位点定向突变发生,pH依赖性研究和基于机制的抑制试验.
主要成果:
- 皮克DH2将d-酒精基质转化为具有可逆动力学 (Keq = 1.22) 的过烯酸产品.
- 证明了严格的基质特异性,对表皮质基质没有转换.
- 确定了一个关键的电离组 (pKa = 7.0) 并描述了基于机制的抑制剂的不可逆转的无活化.
结论:
- 皮克DH2是一个强大的和特定的PKS DH域,适合准备性化学酶合成.
- 对PikDH2的动力学和机械学见解为了解PKS DH功能提供了基础.
- 这项工作促进了PKS路径的设计,以发现新的自然产品.
更多相关视频
09:08From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028
Published on: January 13, 2017
17.9K
11:27X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050
Published on: May 13, 2020
4.4K
相关概念视频
Conservation of Protein Domains Over Different Proteins
15.1K
Protein domains are small structurally independent units that are part of a single amino acid chain. Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
15.1K
Acid-Catalyzed Dehydration of Alcohols to Alkenes
25.6K
In a dehydration reaction, a hydroxyl group in an alcohol is eliminated along with the hydrogen from an adjacent carbon. Here, the products are an alkene and a molecule of water. Dehydration of alcohols is generally achieved by heating in the presence of an acid catalyst. While the dehydration of primary alcohols requires high temperatures and acid concentrations, secondary and tertiary alcohols can lose a water molecule under relatively mild conditions.
25.6K
Photosystem I
72.0K
Although structurally similar to photosystem II (PSII), photosystem I (PSI) is has a different electron supplier and electron acceptor.
Both these photosystems work in concert. An excited electron from PSII is relayed to PSI via an electron transport chain in the thylakoid membrane of the chloroplast, which is comprised of the carrier molecule plastoquinone, the dual-protein cytochrome complex, and plastocyanin. As electrons move between PSII and PSI, they lose energy and must be re-energized...
Both these photosystems work in concert. An excited electron from PSII is relayed to PSI via an electron transport chain in the thylakoid membrane of the chloroplast, which is comprised of the carrier molecule plastoquinone, the dual-protein cytochrome complex, and plastocyanin. As electrons move between PSII and PSI, they lose energy and must be re-energized...
72.0K
ATP Synthase: Structure
17.9K
ATP synthase or ATPase is among the most conserved proteins found in bacteria, mammals, and plants. This enzyme can catalyze a forward reaction in response to the electrochemical gradient, producing ATP from ADP and inorganic phosphate. ATP synthase can also work in a reverse direction by hydrolyzing ATP and generating an electrochemical gradient. Different forms of ATP synthases have evolved special features to meet the specific demands of the cell. Based on their specific feature, ATP...
17.9K
Allosteric Proteins-ATCase
7.0K
Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
7.0K
