| 反相液相色谱-串联质谱法鉴定油菜蜂花粉中的蛋白质及活性肽 |
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| 引用本文： | 郭静,晏嘉泽,郭明,靳艳. 反相液相色谱-串联质谱法鉴定油菜蜂花粉中的蛋白质及活性肽[J]. 色谱, 2014, 32(3): 284-289. DOI: 10.3724/SP.J.1123.2013.12032 |
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| 作者姓名： | 郭静 晏嘉泽 郭明 靳艳 |
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| 作者单位： | 1. 大连大学环境与化学工程学院, 辽宁 大连 116622;2. 中国科学院大连化学物理研究所, 中国科学院分离分析化学重点实验室, 辽宁 大连 116023 |
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| 基金项目： | 东北特色生物资源综合利用项目(2013SYHZ0020) |
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| 摘 要： | 基于鸟枪法蛋白质组学分析方法，使用反相液相色谱-串联质谱（RPLC-MS/MS）系统分析油菜蜂花粉蛋白质的胰蛋白酶酶解产物，结合数据库检索，共鉴定到353条肽段。鉴定到的肽段所归属的蛋白质中有239个蛋白质可检索到其分子生物学功能，主要功能为结合活性、酶活性、运输活性、抑制活性等。根据血管紧张素转化酶（ACE）抑制肽活性与多肽构效之间的关系，从鉴定到的肽段中筛选并适当修饰后得到5条可能具有ACE抑制活性的肽段，化学合成肽段后进行了活性验证。结果表明5条肽段均具有良好的活性，其中肽段AELDIVLALF和LAVNLIPFP表现出较高的ACE抑制活性，半数抑制浓度（IC50）分别为（10.65±0.50）μmol/L和（23.66±1.08）μmol/L。该方法速度快，成本低，大大缩短了鉴定周期，达到了高通量筛选生物活性肽的目的。
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| 关 键 词： | 蛋白质 反相液相色谱-串联质谱 血管紧张素转化酶抑制肽 油菜蜂花粉 |
| 收稿时间： | 2013-12-27 |
Application of reversed-phase liquid chromatography-tandem mass spectrometry in the identification of protein and bioactivity peptides from rape bee pollen |
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| GUO Jing;YAN Jiaze;GUO Ming;JIN Yan. Application of reversed-phase liquid chromatography-tandem mass spectrometry in the identification of protein and bioactivity peptides from rape bee pollen[J]. Chinese journal of chromatography, 2014, 32(3): 284-289. DOI: 10.3724/SP.J.1123.2013.12032 |
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| Authors: | GUO Jing YAN Jiaze GUO Ming JIN Yan |
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| Affiliation: | 1. College of Environmental and Chemical Engineering, Dalian University, Dalian 116622, China;2. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China |
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| Abstract: | Based on the shotgun proteomic method, rape bee pollen protein was prepared with ultrasonic extraction and digested by trypsin, then separated and sequenced by reversed-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), followed by protein database searching. After the above analysis, 353 peptides were identified and the molecular biological functions of 239 proteins have been known. The identified molecular biological functions of these proteins mainly included binding activity, enzyme activity, transporter activity, inhibitor activity and so on. Five peptides were obtained after the screening and appropriate amino acid modification among the identified 353 peptides, according to the relationship between the sequence structure of the bioactivity peptide and angiotensin converting enzyme (ACE) inhibitor activity. The five peptides were speculated to have ACE inhibitor activity and synthesized to detect ACE inhibitor activity. The results showed that all of the five peptides had good ACE inhibitor activity. The peptides of AELDIVLALF and LAVNLIPFP among the five peptides displayed especially good ACE inhibition with half maximal inhibitory concentration (IC50) of (10.65±0.50) μmol/L and (23.66±1.08) μmol/L, respectively. This method is rapid, low-cost and achieves the goal of high-throughput screening of bioactivity peptides that greatly shorten the period of identification compared with traditional methods. |
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| Keywords: | reversed-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) protein angiotensin converting enzyme (ACE) inhibitory peptides rape bee pollen |
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