中文摘要：游离脂肪酸受体1（FFAR1或GPR40）在治疗2型糖尿病方面引起了人们的关注，并开发出多种小分子激动剂。然而，大多数FFAR1激动剂以及内源性配体，如亚油酸，具有高亲脂性，而其高亲脂性与脱靶毒性有关。因此需要关注毒性较小的新配体。本研究筛选了具有FFAR1激动剂活性的多肽作为新的候选配体。首先，使用噬菌体展示鉴定对FFAR1具有高亲和力的多肽；然后通过TGF-α脱落试验评估噬菌体展示测定的多肽的激动剂活性；最后，为了提高肽的FFAR1激动剂活性，进行了包含单氨基酸替换和序列分析。使用120种理化性质进行逻辑回归（LR）分析，预测具有高FFAR1激动剂活性的多肽。噬菌体展示法测定STTGTQY可促进胰腺MIN6细胞葡萄糖刺激的胰岛素分泌。此外，与STTGTQY相比，LR分析预测的STKGTF在低浓度下显示出高胰岛素分泌。可见，多肽可能是新型FFAR1激动剂的候选药物。
外文摘要：Free fatty acid receptor 1 (FFAR1 or GPR40) has attracted attention for the treatment of type 2 diabetes mellitus, and various small-molecule agonists have been developed. However, most FFAR1 agonists as well as endogenous ligands, such as linoleic acids, have high lipophilicity, and their high lipophilicity is related to off-target toxicity. Therefore, we need to focus on new ligand candidates with less toxicity. In this study, we screened peptides with FFAR1 agonist activity as new ligand candidates. First, we used phage display to identify peptides with high affinity to FFAR1. Next, the agonist activities of peptides determined by the phage display were evaluated by the TGF-alpha shedding assay. Finally, to improve the FFAR1 agonist activity of the peptide, we performed an inclusive single amino acid substitution and sequence analysis. Logistic regression (LR) analysis using 120 physiochemical properties was performed to predict peptides with high FFAR1 agonist activity. STTGTQY determined by phage display promoted glucose-stimulated insulin secretion in pancreatic MIN6 cells. Furthermore, STKGTF predicted by the LR analysis showed high insulin secretion at low concentrations compared to STTGTQY. The results of this study suggest that peptides could be new candidates as FFAR1 agonists.
外文关键词：GPR40/FFAR1；Screening；Peptide；Phage display；Machine learning
作者：Yoshioka, K；Yamashita, H；Shimizu, K；Shimomura, S；Shibata, T；Miyazaki, J；Honda, H
作者单位：Nagoya Univ；Osaka Univ
期刊名称：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
期刊影响因子：2.985
出版年份：2021
出版刊次：
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