高效液相色谱/质谱法识别不同明胶酶解产物中特征多肽  

Identification of Specific Peptides in Digested Gelatins by High Performance Liquid Chromatography/Mass Spectrometry

作　　者：张贵锋[1] 刘涛[1] 王前[1] 雷建都[1] 马光辉[1] 苏志国[1] 

机构地区：[1]中国科学院过程工程研究所生化工程国家重点实验室,北京100080

出　　处：《分析化学》2008年第11期1499-1504,共6页Chinese Journal of Analytical Chemistry

基　　金：863计划(No.2007AA021604);973计划(No.2007CB714305)资助项目

摘　　要：在不同动物胶原蛋白序列比对基础上,利用高效液相色谱/质谱联用技术(HPLC/MS)建立一种通过特征多肽进行明胶鉴别的方法。实验以牛明胶和猪明胶为对象,序列比对结果表明,牛和猪的Ⅰ型胶原蛋白α1链和α2链均存在氨基酸序列差异。利用胰蛋白酶将牛明胶和猪明胶降解,通过HPLC/MS分析了两种明胶酶解产物中的多肽片段。结果表明:牛明胶和猪明胶酶解产物中均可检测出存在序列差异的特征性多肽,其氨基酸序列差异与两种胶原蛋白序列比对结果中的序列差异一致;酶解产物中特征多肽上的脯氨酸存在不同程度的羟基化修饰,特征多肽的相对含量主要受明胶分子量范围影响。利用HPLC/MS检测明胶酶解产物中的特征多肽是一种鉴别牛明胶和猪明胶的有效方法。Sequence analysis of bovine and porcine collagen type Ⅰ indicates that some partial sequences are specific for bovine or porcine collagen type Ⅰ , which can be used as marker peptides for gelatin differentiation. Bovine and porcine gelatins were digested with trypsin. The digest mixtures were analyzed with high performance liquid chromatography/mass spectrometry （HPLC/MS）. Marker peptides were identified by the method of peptide sequence tag. The results indicated that the bovine and porcine gelatins could be differentiated according to the sequences of marker peptides. Further analysis indicated that some prolines in the marker peptides were hydroxylated, which should be taken into account to increase the confidence in peptide identification. The relative abundance of marker peptides in gelatin of different molecular weight range was also investigated. The results indicate that identification of the marker peptides in the digested gelatins by HPLC/MS is an alternative method for differentiation of bovine and porcine gelatins.

关 键 词：明胶 酶解 特征多肽 高效液相色谱质谱联用 多肽识别 

分 类 号：Q51-3[生物学—生物化学]