鹿角与鹿茸蛋白质鉴定

Identification of proteins from Cervi Cornu and Cervi Cornu Pantotrichum

目的 鉴定鹿角与鹿茸蛋白质。方法 通过SDS-PAGE、2-DE分析鹿角与鹿茸蛋白质组成,比较两者电泳图谱,找出各自稳定的蛋白质条带或斑点及其差异。将两者SDS-PAGE蛋白质条带以胰蛋白酶进行胶内消化后,采用MALDI-TOF/TOF质谱分析,通过MASCOT检索匹配以鉴定蛋白质。结果 在SDS-PAGE图谱中,鹿角在约75、66.2 kD处呈现出2个清晰的蛋白质条带,而鹿茸则在约75、61、45、20 kD处呈出现4个蛋白质条带;2-DE分析显示,鹿角蛋白质斑点集中于Ⅰ区(pH5~7,M_(w) 45 kD)以及Ⅱ区(pH10,M_(w) 66.2 kD);鹿茸蛋白质斑点则主要分布在Ⅲ区(pH5~7,M_(w) 45 kD),另外,在pH5~6和M_(w) 20 kD的区域内存在3个稳定出现的蛋白质斑点。通过生物质谱分析,分别从鹿角与鹿茸中鉴定出2、4个蛋白质,评分高于100。结论 鹿角与鹿茸蛋白质电泳图谱存在明显差异,可为两者鉴别提供科学依据。

AIM To identify proteins from Cervi Cornu and Cervi Cornu Pantotrichum.METHODS SDS-PAGE and 2-DE were used to compare the protein composition of Cervi Cornu and Cervi Cornu Pantotrichum,and their protein profiles to explore their respective protein bands or spots and ascertain the difference.Subsequently,the SDS-PAGE protein bands subjected to in-gel tryptic digestion had their result ant peptide fragments analyzed by MALDI-TOF/TOF-MS for further identification based on MASCOT matching.RESULTS In SDS-PAGE profiles,two clear protein bands were shown at ca.75 kD and 66.2 kD for Cervi Cornu,and four protein bands were shown at ca.75 kD,61 kD,45 kD and 20 kD for Cervi Cornu Pantotrichum.In 2-DE profiles,Cervi Cornu found its main protein spots distribution in zoneI(pH5-7,M_(w) 45 kD) and zoneⅡ(pH10,M_(w) 66.2 kD);and Cervi Cornu Pantotrichum found its main protein spots distribution in zoneⅢ(pH5-7,M_(w) 45 kD) in addition to three protein spots in area of pH5-6,M_(w) 20 kD.The biological mass spectrum analysis revealed two and four protein composition differences between Cervi Cornu and Cervi Cornu Pantotrichum,presenting higher than 100 scores.CONCLUSION The protein profile differences between Cervi Cornu and Cervi Cornu Pantotrichum provide scientific evidence for their identification.