结构和生化分析显示,酸抑制了DAHP合成酶,DAHP合成酶是石基胺通路中的关键参与者
Kuldeep Jangid1, Jai Krishna Mahto1, K Amith Kumar1
1Department of Biosciences and Bioengineering, IIT Roorkee, Roorkee, India.
Archives of biochemistry and biophysics
|November 20, 2024
概括
昆酸通过向必不可少的石基酸途径酶,3-脱氧-D-阿拉比诺-素酸-7-酸盐合成酶 (DAHPS) 来抑制细菌的生长. 这一发现为开发针对各种细菌感染的新抗菌剂提供了有希望的支架.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 微生物学 微生物学
- 药物发现 药物发现 药物发现
背景情况:
- 石基胺通路对芳香氨基酸生物合成至关重要,在动物中不存在,使其酶成为有吸引力的抗微生物点.
- 3-脱氧-D-氨酸-氨酸-7-酸盐合成酶 (DAHPS) 催化该途径的第一个承诺步骤,是关键的调节点.
- 之前的研究已将酸确定为巴西亚细菌的DAHPS抑制剂,这表明研究其成分,如酸.
研究的目的:
- 为了研究酸 (QA) 作为一种潜在的抑制剂的Providencia alcalifaciens DAHPS (PaDAHPS).
- 为了确定由氨酸 (Phe) 和酸抑制PADAHPS的结构基础.
- 评估酸对各种细菌物种的抗菌活性.
主要方法:
- 确定了对氨敏感的PaDAHPS与Phe和QA复合的晶体结构,分别分辨率为2.5 Å和2.68 Å.
- 进行了生物物理测定,以评估Phe和QA与PaDAHPS的结合亲缘关系.
- 进行了体外抑制研究,以量化QA和Phe对PaDAHPS活性的抑制作用.
- 评估了QA的最小抑制度 (MIC) 与细菌病原体小组相比.
主要成果:
- 晶体结构显示,QA与PaDAHPS上的Phe结合于相同的活性部位.
- 生物物理测试显示,对QA (KD = 42 μM) 和Phe (KD = 32 μM) 的结合亲缘关系相似.
- 实验室研究表明,QA和Phe都抑制了PaDAHPS活性 (QA的Ki = 382μM,Phe的132μM).
- 酸表现出广泛的抗菌活性,抑制了*Pseudomonas aeruginosa*, *Moraxella catarrhalis*, *Providencia alcalifaciens*, *Staphylococcus aureus* 和 *Escherichia coli* (MICs 2.55 mg/ml) 的生长.
结论:
- 酸有效地抑制Providencia alcalifaciens DAHPS,并表现出抗菌性质.
- 结构和生物物理数据为DAHPS受酸抑制的机制提供了洞察力.
- 酸代表了开发针对细菌的新型抗微生物药物的有前途的支架,这些药物针对细菌的shikimate通路.
更多相关视频
08:57Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
Published on: February 24, 2018
10.0K
09:27Functional Complementation Analysis FCA: A Laboratory Exercise Designed and Implemented to Supplement the Teaching of Biochemical Pathways
Published on: June 24, 2016
17.6K
相关概念视频
01:30The Electron Transport Chain
16.1K
The electron transport chain or oxidative phosphorylation is an exothermic process in which free energy released during electron transfer reactions is coupled to ATP synthesis. This process is a significant source of energy in aerobic cells, and therefore inhibitors of the electron transport chain can be detrimental to the cell's metabolic processes.
Inhibitors of the electron transport chain
Rotenone, a widely used pesticide, prevents electron transfer from Fe-S cluster to ubiquinone or Q...
Inhibitors of the electron transport chain
Rotenone, a widely used pesticide, prevents electron transfer from Fe-S cluster to ubiquinone or Q...
16.1K
01:19Allosteric Proteins-ATCase
5.7K
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...
5.7K
01:28Biosynthesis of Nucleic Acids
7
Nucleic acid biosynthesis is a fundamental biochemical process that produces the purine and pyrimidine nucleotides essential for DNA and RNA synthesis. This pathway maintains a balanced nucleotide pool, preventing imbalances that could jeopardize genetic integrity and cellular function. Given the crucial role of nucleotides, their synthesis is tightly regulated to ensure proper cellular homeostasis.Purine BiosynthesisThe biosynthesis of purine nucleotides begins with ribose-5-phosphate, a...
7
01:11Aryldiazonium Salts to Azo Dyes: Diazo Coupling
2.9K
The reaction of weakly electrophilic aryldiazonium (also called arenediazonium) salts with highly activated aromatic compounds leads to the formation of products with an —N=N— link, called an azo linkage. This reaction, presented in Figure 1, is known as diazo coupling and occurs without the loss of the nitrogen atoms of the aryldiazonium salt. Highly activated aromatic compounds such as phenols or arylamines favor the diazo coupling reaction. The coupling generally occurs at the...
2.9K
01:37The Citric Acid Cycle: Overview
16.1K
In aerobic organisms, the citric acid cycle is the second stage of cellular respiration wherein molecules derived from the breakdown of carbohydrates, proteins, and fats are oxidized into carbon dioxide and energy. This process is also known as the tricarboxylic acid (TCA) cycle as the first product of the cycle, citric acid, contains three carboxyl groups in its structure. Alternatively, this cycle is also referred to as the Krebs cycle, in honor of its discoverer Sir Hans Krebs.
The citric...
The citric...
16.1K
01:28The Citric Acid Cycle: Output
7.7K
The citric acid cycle is termed an amphibolic pathway as it operates both anabolically and catabolically. The cyclic reactions balance the flux of the substrates to provide an optimal concentration of NADH and ATP to the cell.
Regulation of Citric Acid Cycle
The citric acid cycle is regulated in several ways, including feedback inhibition, regulation of enzyme activities, and associated anaplerotic or cataplerotic pathways.
The primary substrate of the TCA cycle—acetyl CoA—is...
Regulation of Citric Acid Cycle
The citric acid cycle is regulated in several ways, including feedback inhibition, regulation of enzyme activities, and associated anaplerotic or cataplerotic pathways.
The primary substrate of the TCA cycle—acetyl CoA—is...
7.7K