摘要
利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分离了银杏种子中的糖蛋白组分,进一步用氨水催化释放N-糖链,并采用电喷雾离子质谱(ESI-MS)及在线液相色谱-质谱联用(LC-UV-MS/MS^2)等技术对胶条上释放的N-糖链进行了定性定量分析.结果表明,从银杏种子中分离得到11种糖蛋白,共检测到11条N-糖链,包括高甘露糖型(4. 88%)和复杂型(95. 12%) 2种类型,其中分子量为21000,36000和50000的糖蛋白所释放的核心α-1,3-岩藻糖和β-1,2-木糖修饰的N-糖链所占比率较高,分别为68. 23%,64. 37%和75. 09%.本研究对进一步研究银杏糖蛋白功能具有重要意义.
The glycoproteins in Ginkgo seeds were separated and isolated by sodium dodecyl sulfate polyacry-lamide gel electrophoresis(SDS-PAGE).The N-glycans released from glycoproteins were qualitatively and quantitatively analyzed by electrospray ionization mass spectrometry(ESI-MS)and on-line liquid chromatography-mass spectrometry(LC-UV-MS/MS 2).As a result,eleven glycoproteins containing oligomannose type glycans(4.88%)and complex type glycans(95.12%)were obtained.And most of the N-glycans were modified by the coreα-1,3-andβ-1,2-xylose in the 21000,36000 and 50000 glycoproteins,which accounted for 68.23%,64.37%and 75.09%,respectively.This study can highlight the understanding of protein glycosylation in Ginkgo and provide foundation in the research of Ginkgo biloba seed allergens.
作者
李成
王承健
晋万军
韩健利
杨梅芳
郜茜
黄琳娟
王仲孚
LI Cheng;WANG Chengjian;JIN Wanjun;HAN Jianli;YANG Meifang;GAO Xi;HUANG Linjuan;WANG Zhongfu(College of Life Sciences,Northwest University,Xi'an 710069,China;College of Food Sciences and Technology,Northwest University,Xi’an 710069,China)
出处
《高等学校化学学报》
SCIE
EI
CAS
CSCD
北大核心
2019年第1期69-75,共7页
Chemical Journal of Chinese Universities
基金
国家自然科学基金(批准号:31670808
31600647)
陕西省教育厅自然科学专项项目(批准号:16JK1782)
西北大学校内基金(批准号:15NW18)资助~~
关键词
银杏糖蛋白
十二烷基硫酸钠聚丙烯酰胺凝胶电泳
N-糖苷酶F
氨水催化
电喷雾离子质谱
Ginkgo glycoprotein
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
PNGase F
Aqueous ammonia
Electrospray ionization mass spectrometry
