作　　者： ; ; ; ; ; ;

机构地区： 吉林大学公共卫生学院

出　　处： 《分析化学》 2009年第9期1269-1274,共6页

摘　　要： 采用高效液相色谱(HPLC)SCX及C4色谱柱从人乳样品的乳清部分分离与纯化出骨桥蛋白(Osteopontin,OPN),在一定条件下将其用胰蛋白酶酶解后,利用高分辨傅立叶变换离子回旋共振质谱(FT-ICR-MS),结合微喷雾(Nano-spray)技术,并借助Mascot软件数据库,获得了OPN相关多肽片段GDSVVYGLR和QNLLAPQTLPSK的质谱结构信息,分析了其裂解机理。研究表明,利用FT-ICR-MS并结合HPLC等化学方法,可以高效、快速地分离与纯化人乳样品中的OPN,借助Mascot软件数据库进行质谱分析,能够简洁、准确地鉴定相关多肽片段的结构信息,为OPN多肽片段不同翻译后修饰位点的确定与其生物学活性关系的研究提供了理论依据。 Fourier transform-ion cyclotron resonance-mass spectrometric（FT-ICR-MS） technique has the significant advantages in sensitivity and dynamic range for the detection of different fragmentations of peptides and proteins at lower levels.The osteopontin（OPN） was isolated and purified from the healthy human milk using HPLC by SCX and different types of C4 columns combined with FT-ICR-MS and Mascot software search.After digesting the purified OPN sample by the specific enzyme,some correlative mass structural information of the polypeptide fragmentations such as GDSVVYGLR and QNLLAPQTLPSK was identified by nano-spray HPLC and FT-ICR-MS.It shows that nano-spray HPLC combined with FT-ICR-MS and Mascot search can be used as a high efficient method to purify and isolate OPN from the human milk.The study will provide more information of the different modifications on its related polypeptides and the relationship between its structure and bioactivity.

关 键 词： 傅立叶变换离子回旋共振质谱 人乳 骨桥蛋白 多肽片段

领　　域： [理学] [理学] [核科学技术]