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基于生物信息学和质谱技术的阿胶特异性肽段筛选与鉴定

投稿时间：2019-03-20 点此下载全文

引用本文：王芳,范雨,叶茂,曹秀君,童芯锌,彭成,国锦琳.基于生物信息学和质谱技术的阿胶特异性肽段筛选与鉴定[J].中国现代中药,2019,21(9):1256-1261

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作者中文名	作者英文名	单位中文名	单位英文名	E-Mail
王芳	WANG Fang	成都中医药大学中药资源系统研究与开发利用重点实验室中药材标准化重点实验室，四川成都611137	The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine，Key Laboratory of Systematic Research，Development and Utilization of Chinese Medicine Resources Key Laboratory Breeding Base，Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China
范雨	FAN Yu	成都中医药大学中药资源系统研究与开发利用重点实验室中药材标准化重点实验室，四川成都611137	The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine，Key Laboratory of Systematic Research，Development and Utilization of Chinese Medicine Resources Key Laboratory Breeding Base，Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China
叶茂	YE Mao	成都中医药大学中药资源系统研究与开发利用重点实验室中药材标准化重点实验室，四川成都611137	The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine，Key Laboratory of Systematic Research，Development and Utilization of Chinese Medicine Resources Key Laboratory Breeding Base，Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China
曹秀君	CAO Xiu-jun	成都中医药大学中药资源系统研究与开发利用重点实验室中药材标准化重点实验室，四川成都611137	The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine，Key Laboratory of Systematic Research，Development and Utilization of Chinese Medicine Resources Key Laboratory Breeding Base，Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China
童芯锌	TONG Xin-xin	成都中医药大学中药资源系统研究与开发利用重点实验室中药材标准化重点实验室，四川成都611137	The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine，Key Laboratory of Systematic Research，Development and Utilization of Chinese Medicine Resources Key Laboratory Breeding Base，Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China
彭成	PENG Cheng	成都中医药大学中药资源系统研究与开发利用重点实验室中药材标准化重点实验室，四川成都611137	The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine，Key Laboratory of Systematic Research，Development and Utilization of Chinese Medicine Resources Key Laboratory Breeding Base，Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China
国锦琳	GUO Jin-lin	成都中医药大学中药资源系统研究与开发利用重点实验室中药材标准化重点实验室，四川成都611137	The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine，Key Laboratory of Systematic Research，Development and Utilization of Chinese Medicine Resources Key Laboratory Breeding Base，Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China	国锦琳，教授，研究方向：名贵中药资源与鉴定研究；E-mail：guo596@163.com

基金项目:四川省青年科技创新团队项目（19CXTD0055）

中文摘要:目的：建立基于生物信息技术和液质联用技术识别特征肽鉴别阿胶的新方法。方法：利用生物信息技术对驴皮、牛皮和猪皮I型胶原蛋白α1链氨基酸序列进行同源性比对发现3种皮类胶原蛋白氨基酸序列存在差异，通过生物学软件模拟胰酶酶解，分析比对驴皮、牛皮、猪皮来源I型胶原蛋白α1链酶解肽段，然后通过实验对样品前处理条件进行系统摸索，采用超高效液相色谱-四级杆飞行时间质谱（UPLC/Q-TOF-MS）进行验证。结果：筛选出12个驴皮的特异性肽段，鉴定出1个与预测结果一致的驴皮特异性肽段，其质荷比（m/z）为766.4，分子量为1 530.802 2，对应的多肽序列为GEAGPAGPAGPIGPVGAR，利用特征肽段对10组阿胶样品进行鉴定。结论：生物信息技术结合UPLC/Q-TOF-MS检测出的驴皮特征肽可有效、特异、准确地对阿胶药材进行专属性鉴定。

中文关键词:阿胶鉴定胰酶特异肽段生物信息学超高效液相色谱-四级杆飞行时间质谱

Screening and Identification of Characteristic Peptides of Donkey-hide Gelatin Based on Bio-informatics Technique and Mass Spectrometry

Abstract:Objective：To establish a new method based on biological information technology and liquid chromatography mass spectrometry technology to identify characteristic peptides for the identification of E-jiao（donkey-hide gelatin）.Methods：Homologous comparison of the α1 chain amino acid sequences of type I collagen from donkey skin，cow skin and pig skin using bioinformatics technology showed that there were differences in amino acid sequences of three kinds of skin collagen.The trypsin digest was simulated by biological software，and the enzymolysis peptides of type I collagen from donkey skin，cow skin and pig skin were compared.Then，the sample pretreatment conditions were systematically explored through experiments，and ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometer（UPLC/Q-TOF-MS）for verification.Results：Twelve specific peptides of donkey-hide were screened out，and one donkey specific peptides was identified in accordance with the predicted results with a mass-to-charge ratio（m/z）of 766.4，mass of 1 530.802 2，corresponding peptide sequence of GEAGPAGPAGPIGPVGAR.Ten groups of donkey-hide gelatin samples were identified by using characteristic peptides.Conclusion：The detection characteristic peptide in the digested gelatins using UPLC/Q-TOF-MS coupled with bio-informatics technique could be used to identify E-jiao effectively，specifically and accurately.

keywords:E-jiao identification characteristic peptides trypsin enzyme digestion UPLC/Q-TOF-MS bio-informatics technique

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