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基于激光解析/离子化-飞行时间质谱技术的中药阿胶蛋白质组分析 预览 被引量:20

Analysis of the traditional Chinese medicine Donkey-hide gelatin proteome based on surface enhanced laser desorption/ionization time-of-flight mass spectrometry
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摘要 目的:应用激光解析,离子化-飞行时间质谱技术对传统中药阿胶蛋白/肽成分进行蛋白质组初步分析,建立阿胶蛋白质质量指纹图。方法:实验于2004-06/2005—03在湖南中医药大学蛋白质组学实验室完成。采用饱和硫酸胺沉淀,透析脱盐冻干法获得阿胶水溶性蛋白/肽,采用Ciphergen Biosysterns Inc.ProteinChip Biology System(美国PBSⅡ+)及配套的NP10芯片、激光解析,离子化-飞行时间质谱技术,分析阿胶Mr 1500~13000区间蛋白质、肽分布及其相对分子质量。结果:M2000-4000区间显示4个相对分子质量峰,可能是2种肽;Mr 4000-5000区间显示8个相对分子质量峰,可能为2种肽;Mr 5000-5500区间显示16个相对分子质量峰,约为3种肽;Mr 5500-6200区间无峰;Mr8100-8200区间显示5个相对分子质量峰,可能为1种肽;Mr 11200-11400区间显示5个相对分子质量峰,可能为1种蛋白质。不同质量浓度稳定获得的有意义蛋白/肽共计9个。不同相对分子质量肽之间差分别提示可能为丙氨酸,丝氨酸、苯丙氨酸残基相对分子质量。结论:通过分析,可以形成阿胶蛋白质/肽成分质量指纹图,可作为阿胶数字化质控标准;并为进一步分离、纯化及验证阿胶功能相关活性蛋白质/肽组分提供可靠信息。 AIM: To set up the proteomic fingerprinting of Donkey-hide gelatin by using surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). METHODS: The experiment was performed at the Laboratory of Proteome, Hunan University of Traditional Chinese Medicine from June 2004 to March 2005. By means of dialysis-desalination-freeze drying with satisfying sulphuric acid amine sediment, the water soluble protein/ peptide from the Donkey-hide gelatin was gotten. Ciphergen Biosystems Inc.ProteinChip Biology System (America PBS Ⅱ +) and combined NP10array, SELDI-TOF-MS were adopted to analyze the Donkey-hide gelatin's Mr 1 500-13 000 Da protein, peptide distribution and its relative molecular mass (RMM). RESULTS:There were 4 peaks, disl01aying 2 kinds of peptide between Mr 2 000-4 000 Da; 8 peaks, displaying 2 kinds of peptide between Mr 4 000-5 000 Da; 16 peaks, about 3 kinds of peptide between Mr 5 000-5 500 Da; no peak between Mr 5 500-6 200 Da; 5 peaks, belonged to a kind of peptide between Mr 8 100-8 200 Da; and 5 peaks, belonged to a kind of peptide between Mr 11 200-11 400 Da. There were 9 kinds of meaningful protein/peptide with different mass concentrations. It might display the lactamine, serine, phenyialanine residues from the disparities of different RMM. CONCLUSION: The Donkey-hide gelatin protein/peptide mass fingerprinting (PMF) can be obtained as digital quality control standard of the Donkey-hide gelatin. Reliable information on further isolation, purification and verification of the Donkey-hide gelatin protein/peptide is provided.
作者 王若光 尤昭玲 刘小丽 刘惠萍 秦明春 李春梅 胡维新 Wang RG, You ZL, Liu XL, Liu HP, Qin MC, Li CM, Hu WX(1.College of Integrated Traditional and Western Medicine, Hunan University of Traditional Chinese Medicine, Changsha 410007, Hunan Province, China; 2.Biological Science and Technology College, Central South University, Changsha 410007, Hunan Province, China)
出处 《中国组织工程研究与临床康复》 CAS CSCD 北大核心 2007年第13期 2518-2521,共4页 Journal of Clinical Rehabilitative Tissue Engineering Research
基金 湖南省自然科学基金项目资助(03JJY3042) 湖南省社会发展重大项目资助(03SSY1012).
作者简介 王若光☆,男,1967年生,河南省遂平县人,汉族,2003年湖南中医药大学毕业,博士,教授。主要从事中药成分分子药理学的研究。dplnw@public.cs.hn.cn
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