通过浮躁的黄素依赖酶进行基化
Andrew J Rice1,2, Mayuresh G Gadgil2, Paola Bisignano3
1Department of Biochemistry, Vanderbilt University School of Medicine-Basic Science, Department of Chemistry, Vanderbilt University, Nashville, TN, 37232, USA.
Angewandte Chemie (International ed. in English)
|August 30, 2025
概括
研究人员发现了一种多功能酶,ChlH (黄素依赖酶),它可以修改中的内部氨基酸. 这一发现扩大了生物技术中的酶合成和修饰的潜力.
科学领域:
- 生物化学
- 酵素学
- 合成生物学
背景情况:
- 氨基酸的酶修饰是至关重要的,但通常受到酶特异性的限制.
- 黄素依赖酶 (FDHs) 通常会修改特定的或末.
- 已知FDH的狭窄基质范围和辅助因子要求限制了它们的合成应用.
研究的目的:
- 描述ChlH,一种来自氨酸生物合成基因群的新型氨酸酶.
- 调查ChlH的基质范围和催化活性.
- 探索ChlH在广泛和蛋白质修饰方面的潜力.
主要方法:
- 对ChlH的酶表征
- 基质ChlA的扫描突变发生.
- 进行分子动力学模拟.
- 在各种基质上进行化测试,包括RiPP和药理相关的.
- 没有细胞的生物合成试验.
主要成果:
- 与其他FDH不同的是,除了N和C终端的Trp外,ChlH还有效地化酸 (Trp) 的内部残留物.
- 突变和模拟显示了基质识别和特异性的洞察力,解释了原生氨酸生物合成的局限性.
- 野生型ChlH证明了广泛的适用性,成功地化了各种和蛋白质基质.
- 无细胞测试阐明了ChlH的基质偏好,突出了它的杂乱性.
结论:
- 在FDH中,ChlH表现出独特和广泛的基质特异性,使其能够在多种序列中修改内部的Trp残留物.
- ChlH的杂交性表明其在修改广泛的和蛋白质基质方面的应用潜力很大.
- 这种酶代表了合成生物学和生物技术的宝贵工具,促进了新的修饰.
相关概念视频
Repressible Operon: trp Operon
135
The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...
135
Transcription Attenuation in Prokaryotes
16.0K
Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure. Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
16.0K
Amino Acid Biosynthetic Pathways
131
Amino acid biosynthesis is essential for cell growth, protein synthesis, and metabolic regulation. Cells generate essential and non-essential amino acids from metabolic intermediates to sustain vital biological functions. These intermediates originate from key metabolic pathways: glycolysis, the tricarboxylic acid (TCA) cycle, and the pentose phosphate pathway. Important precursors include α-ketoglutarate, pyruvate, oxaloacetate, phosphoenolpyruvate, and erythrose-4-phosphate, which...
131
Protein Transport to the Stroma
1.9K
Chloroplasts are triple membrane structures with an outer membrane, an inner membrane, and a thylakoid membrane, each containing distinct metabolite transporters, membrane translocons, and enzymes. Appropriate sorting and translocating these proteins to their correct membrane systems is essential for chloroplast function.
Protein complexes called the translocon of the outer chloroplast membrane or TOC complex, and the translocon of the inner chloroplast membrane or TIC complex mediate the...
Protein complexes called the translocon of the outer chloroplast membrane or TOC complex, and the translocon of the inner chloroplast membrane or TIC complex mediate the...
1.9K
Translocation of Proteins into the Mitochondria
3.2K
Mitochondrial precursors are translocated to the internal subcompartments via independent mechanisms involving distinct protein machineries called translocases.
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
3.2K
Allosteric Proteins-ATCase
5.9K
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...
5.9K


![Radiosynthesis of 1-2-[18F]Fluoroethyl-L-Tryptophan using a One-pot, Two-step Protocol](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F63025.jpg&w=3840&q=50)