焦虑症在全球范围内流行,极大降低了患者的生活质量。γ-氨基丁酸转氨酶(γ-aminobutyric acid transaminase,GABA-T)作为降解抑制性神经递质γ-氨基丁酸(γ-aminobutyric acid,GABA)的限速酶,抑制其活性可有效缓解焦虑。该研究旨在借助计算机模拟技术,从螺旋藻藻蓝蛋白中筛选出具有抗焦虑潜力的GABA-T抑制肽。研究构建了多层级筛选策略。首先,对藻蓝蛋白氨基酸序列进行虚拟酶切,经PeptideRanker模型筛选出高活性多肽,接着选择分子对接结合能较高的多肽进行毒性、致敏性、水溶性预测,及药物代谢动力学分析,最后通过分子对接和分子动力学模拟揭示候选肽的作用模式。结果表明,酶切获得49条高活性肽,经分子对接选择30条进行理化性质和药代动力学分析,最终可获得3条无毒、无致敏、水溶性和成药性良好的多肽序列CLNGLR、GRF、QGRF。对接结果和动力学模拟揭示,3条多肽可以与GABA-T的关键残基结合,构象在动力学模拟过程中稳定。该研究表明螺旋藻源多肽可以通过抑制GABA降解发挥抗焦虑作用,具备开发为天然抗焦虑药物的潜力。
Anxiety Disorder is prevalent worldwide and significantly impairs patients’ quality of life.γ-aminobutyric acid transaminase (GABA-T) is the rate-limiting enzyme for degrading the inhibitory neurotransmitter γ-aminobutyric acid (GABA), and inhibiting its activity can effectively relieve anxiety.This study aims to screen GABA-T inhibitory peptides with anti-anxiety potential from Spirulina phycocyanin by using computer simulation techniques.A multi-level screening strategy was constructed in this study.Firstly, virtual enzymatic digestion was performed on the phycocyanin amino acid sequence, and highly bioactive peptides were screened out through the PeptideRanker model.Then, polypeptides with high molecular docking binding energy were selected for toxicity, allergenicity, and water solubility predictions, followed by pharmacokinetic analysis.Finally, the mode of action of candidate peptides was revealed through molecular docking and molecular dynamics simulation.The results show that 49 highly bioactive peptides were obtained by enzymatic digestion.After molecular docking, 30 polypeptides with high docking binding energy were selected for physicochemical properties and pharmacokinetic analysis.Finally, three non-toxic, non-allergenic, water-soluble, and drug-like polypeptide sequences (CLNGLR, GRF, and QGRF) were obtained.The docking and dynamics simulation results revealed that these three polypeptides could bind to the key residues of GABA-T and maintain stable conformations during the dynamics simulation.This study indicates that Spirulina-derived polypeptides exhibit anti-anxiety activity by inhibiting GABA degradation, holding potential for development as natural anti-anxiety agents.
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