通过深度学习优化的氨基酸-tRNA合成酶酶表现得像一个准物种
Sourav Kumar Patra1, Nicholas Randolph1, Brian Kuhlman
1Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7260, USA.
Structural dynamics (Melville, N.Y.)
|April 28, 2025
概括
深度学习重新设计了urzymes,改善了研究早期遗传编码的可溶性. 这些工程蛋白质提供了关于祖先蛋白质化学和进化的见解.
科学领域:
- 蛋白质设计和计算生物学.
- 遗传编码和祖先蛋白质的起源.
背景情况:
- 尿酶是氨基酸-tRNA合成酶的活性片段,对于理解早期遗传编码至关重要.
- 之前使用罗塞塔的urzyme设计具有有限的溶解性,阻碍了结构生物学研究.
研究的目的:
- 通过深度学习重新设计urzymes,以提高溶解度和稳定性.
- 研究蛋白质序列,结构,可溶性和催化活性之间的关系.
主要方法:
- 使用深度学习算法ProteinMPNN和AlphaFold2进行urzyme重新设计.
- 使用主要成分分析进行测试,选择了八种优化的 LeuAC 尿酶变体.
主要成果:
- 与原始设计相比,重新设计的urzymes表现出明显改善的溶解度.
- 测试的变种显示了一系列的催化能力和氨基酸特异性.
- 数据允许对促进溶解性和特异性的因素进行统计分析.
结论:
- 深度学习成功地解决了urzyme设计中的可溶性和稳定性限制.
- 重新设计的urzymes为详细研究祖先蛋白质化学提供了一个平台.
- 这些变体为研究诸如蛋白质种群中的自然选择等进化原理提供了一个模型.
相关概念视频
tRNA Activation
18.7K
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
18.7K
Improving Translational Accuracy
8.5K
Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
8.5K
Transfer RNA Synthesis
11.7K
One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
11.7K
Allosteric Proteins-ATCase
5.6K
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...
5.6K
Leaky Scanning
5.0K
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.0K
Catalytically Perfect Enzymes
3.8K
The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
Most enzymes...
Most enzymes...
3.8K


