中国组织工程研究2024，Vol.28Issue(17)：2675-2681,7.DOI:10.12307/2024.445
青蛙皮肤活性肽QUB2984的重组表达及体外活性评价
Recombinant expression and in vitro activity identification of a bioactive peptide QUB2984 from skin secretion of Agalychnis callidryas
唐子琰 1顾顺强 2陈小玲 3王蕾 3马成帮 3周玫 3陈天豹 3杜丽娜 1金义光1
作者信息
- 1. 军事科学院军事医学研究院辐射医学研究所,北京市 100850
- 2. 武警宁夏总队医院急诊科,宁夏回族自治区 银川市 750000
- 3. 英国贝尔法斯特女王大学药学院天然药物研究组,英国贝尔法斯特 BT7 1NN
- 折叠
摘要
Abstract
BACKGROUND:Frog active peptides have rich activities,such as antibacterial and anti-tumor,and are expected to solve the problem of antibiotic resistance.
OBJECTIVE:The active peptide QUB2984 was discovered in the skin secretions of Agalychnis callidryas.Its structure and properties were simulated by bioinformatics.The peptide was synthesized,purified,and identified and its biological functions were investigated.
METHODS:Agalychnis callidryas skin secretions were collected by electrostimulation.The sequence of QUB2984 was obtained through constructing a cDNA library with isolated mRNA.BLAST was used for peptide sequence alignment.Besides that,Iterative Threading ASSEmbly Refinement(I-TASSER)and HeliQuest tools were used for protein secondary structure simulation.It was synthesized by solid-phase peptide synthesis,purified by reverse-phase high-performance liquid chromatography,and structurally confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.The purified peptide was used to evaluate its biological activity.Its antibacterial effect was evaluated by the minimum inhibitory concentration method.Its cytotoxic effect was detected by MTT assay.Its safety was investigated by a hemolysis test.
RESULTS AND CONCLUSION:(1)Peptide QUB2984 had basically α-spiral structure,with a relatively intact hydrophobic surface,and a certain destructive ability to biofilm.The third amino acid position of QUB2984 was composed of W and had a G-X-G structure.(2)The minimum inhibitory concentration of QUB2984 against gram-positive Staphylococcus aureus was 2 μmol/L,the minimum inhibitory concentration against gram-negative Escherichia coli was 2 μmol/L,and the minimum inhibitory concentration against the fungus Candida albicans was 8 μmol/L.(3)The active peptide QUB2984 had obvious inhibitory effect on human non-small cell lung cancer cells NCI-H838 at 10-5 mol/L concentration,and the hemolytic effect on horse red cells at 64 μmol/L concentration was 50%.(4)The results showed that QUB2984 had anti-bacterial and anti-cancer activity,and it had a positive charge of +3,which was conducive to contact with bacteria or cells.关键词
生物活性肽/生物合成/结构模拟/构效关系/分离纯化/抗菌/抑癌/溶血评价Key words
bioactive peptide/biosynthesis/structural simulation/structure-activity relationship/isolation and purification/antibacterial/anticancer/hemolysis evaluation分类
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唐子琰,顾顺强,陈小玲,王蕾,马成帮,周玫,陈天豹,杜丽娜,金义光..青蛙皮肤活性肽QUB2984的重组表达及体外活性评价[J].中国组织工程研究,2024,28(17):2675-2681,7.
