基于激光解析/离子化飞行时间质谱技术的中药龟甲胶蛋白质组分析

Analysis of Chinese medicine colla carapacis et plastri testudinis proteome with technique SELDI-TOF

目的对传统中药龟甲胶蛋白质、肽成分进行组蛋白质组分析。方法采用饱和硫酸胺沉淀,透析脱盐法获得龟甲胶蛋白质成分,美国Ciphergen Biosystems Inc.ProteinChip Biology System(PBSⅡ+)及配套的NP10芯片、激光解析/离子化飞行时间质谱(SELDI-TOF MS,Surface enhanced laser desorption/ionization time-of-flightmass spectro metry)技术,分析了龟甲胶1 500～13 000 Da区间蛋白质、肽分布及其分子量。结果龟甲胶蛋白质/肽质量1 500～3 000区间显示1个分子量峰;4 800～5 400显示3个分子量峰,其中相对分子量分别相差410.8、151.7;8 000～8 600区间显示1个分子量峰;10 000～13 000区间显示1个分子量峰。共获得6个肽分子量峰。结论通过分析,可以形成龟甲胶蛋白质/肽成分质量指纹图,可作为龟甲胶数字化质控标准;并为进一步分离、纯化及验证龟甲胶功能相关活性蛋白质/肽成分打下基础。

Objective To analyze the proteome and peptide in traditional Chinese medicine colla carapacis et plastri testudinis. Methods The colla carapacis et plastri testudinis protein/peptide By means of dialysis-desalination-freeze drying with satisfying sulphuric acid amine sediment and analyze the peptide＇s M-interval 1500-13000Da protein, peptide distribution and its Molecular Weight with Ciphergen Biosystems Inc. ProteinChip Biology System （American PBSⅡ ＋） and the combined NP10array, SELDI-TOF-MS, Surface enhanced laser desorption/ionization time-of-flight mass spectrometry）technique. Results The 6 table humps of protein and peptide Mol Wt.Among were observed, in which there was 1 hump of Mol Wt between the interval 1500-3000Da, the different relative Mol Wt were 410.8 and 151.7.;one hump of Mol Wt between the interval 8000-8600 Da, one hump of Mol Wt between the inter val 10000-13000Da, and were 3 humps of Mol Wt between the interval 4800-5400Da. Conclusion The fingerprinting of the colla carapacis et plastri testudinis protein and peptide Mol Wt can be measured, which can be the digitizated quality control standard of the Donkey-hide gelatin, and works as the foundation to further separating, purifying and detecting the colla carapacis et plastri testudinis protein and peptide.