摘要
建立云南与四川僵蚕药材的总蛋白质指纹图谱及分级指纹图谱,对比分析以发现其中的差异表达蛋白质分子并进行鉴定及生物信息学分析,探索其用于产地鉴定的可行性。提取两种僵蚕的酸溶、碱溶、水溶及醇溶性总蛋白质,对其进行丙酮分级沉淀,对所得样品进行SDS-PAGE指纹图谱对比分析,对所得差异条带进行LC-MS/MS分析,数据库检索后进行生物信息学分析。两种僵蚕的总蛋白质指纹图谱相似度很高,难以据此进行产地鉴定;两种僵蚕的分级指纹图谱则有较显著差异,有望用于产地鉴定;依据分级指纹图谱差异共鉴定出β-葡萄糖苷酶、抗胰凝乳蛋白酶、抗胰蛋白酶、含脂肪酶结构域蛋白等273种蛋白质。云南与四川僵蚕的蛋白质组成及其分级指纹图谱存在显著差异,有望用作产地鉴定的分子依据;醇溶蛋白质构建分级指纹图谱可以提供更加丰富的信息及更好的分辨率;分级指纹图谱技术具有简便易行、分辨率较高的优点,为其他中药材的产地鉴定研究提供了方法学思路。
To establish and compare the total protein fingerprints and the fractional fingerprints of Bombyx Batryticatus from Yunnan and Sichuan,and to discover the differentially expressed protein molecules,and to identify and analyze them by mass spectrum and bioinformatics,to explore the feasibility of its application in geographical origin identification.The total acid-soluble,alkali-soluble,water-soluble and alcohol-soluble proteins of the two kinds of Bombyx mori were extracted,and they were fractional precipitated with acetone.The SDS-PAGE fingerprints of the obtained samples were compared and analyzed.The differential bands were analyzed by LC-MS/MS,and the bioinformatics analysis was conducted after database retrieval.The similarity of the total protein fingerprints of these two kinds of Bombyx Batryticatus was very high,so it is difficult to use for geographical origin identification of this Chinese medicinal material.The fractional fingerprints of these two kinds of Bombyx Batryticatus were relatively different,which is expected to be used for the identification of its geographical origin.According to the difference of fractional fingerprints,273 proteins including chymotrypsin inhibitor,trypsin inhibitor,fumarylacetoacetate hydrolase domain-containing protein and lipase domain-containing protein were identified.There were significant differences in protein composition and fractional fingerprints between Bombyx Batryticatus from Yunnan and Sichuan,which were expected to be used as molecular basis for geographical origin identification.The construction of fractional fingerprints of alcohol-soluble proteins can provide more abundant information and better resolution.The fractional fingerprint technology has the advantages of simplicity and high resolution,which provides a methodological idea for the identification of other Chinese medicinal materials.
作者
宋美营
王立勇
王巧宇
张林松
徐卫东
汤建
闻崇炜
SONG Mei-ying;WANG Li-yong;WANG Qiao-yu;ZHANG Lin-song;XU Wei-dong;TANG Jian;WEN Chong-wei(School of Pharmacy,Jiangsu University,Zhenjiang 212013,China;Zhenjiang Food and Drug Supervision and Inspection Center,Zhenjiang 212013,China;Traditional Chinese Medicine College,Bozhou University,Bozhou 236800,China)
出处
《天然产物研究与开发》
CAS
CSCD
2023年第9期1486-1494,共9页
Natural Product Research and Development
基金
中央本级重大增减支项目(2060302)
江苏省镇江市重点研发计划(社会发展)(SH2018023,SH2021010)
中药原料产品研发安徽普通高校重点实验室(亳州学院)建设项目(KLAHEI18032)。
关键词
僵蚕
蛋白质
分级指纹图谱
产地鉴定
Bombyx Batryticatus
proteins
fractional fingerprints
geographical origin identification
