蕲蛇Ⅱ型胶原蛋白的提取和鉴定表征  

Extraction,purification and identification of type Ⅱ collagen from Agkistrodon acutus

作　　者：谷恒存[1] 胡金波[1] 丁志山[1] 范永升[1] 丁兴红[1] 

机构地区：[1]浙江中医药大学,浙江杭州310053

出　　处：《中国中药杂志》2013年第21期3672-3675,共4页China Journal of Chinese Materia Medica

基　　金：浙江省重点科技创新团队项目(2011R09042-03)

摘　　要：该研究通过限制性胃蛋白酶酶解法提取蕲蛇中Ⅱ型胶原蛋白（type Ⅱ collagen,CⅡ）,并采用离子交换色谱法对其进行纯化,所得蛋白通过SDS-PAGE凝胶电泳、紫外分光光度法、红外吸收光谱法和质谱法进行表征鉴定,结果显示,蕲蛇CⅡ相对分子质量约为130 kDa,其紫外吸收峰约为223 nm;红外图谱中在1 200~1 450 cm-1具有特征吸收,表明蕲蛇CⅡ中有特征序列Gly-X-Y肽段,存在三股螺旋结构。通过对蕲蛇CⅡ与牛CⅡ进行蛋白质肽段质谱图分析对照后发现,两者具有相似肽段和CⅡ结构特征。以上鉴定表征结果证明采用酶解法提取蛋白为蕲蛇CⅡ,可为蕲蛇药材CⅡ的进一步药理研究提供实验基础。The object of the research was to extract, purify and identify the type Ⅱ collagen of Agkistrodon acutus. Type Ⅱ collagen of A. acutus was extracted by enzyme decomposition method, and purified by ion exchange column chromatography. It was characterized by SDS-PAGE gel electrophoresis, ultraviolet spectrophotometry, infrared absorption spectroscopy and mass spectroscopy. The results showed that the size of CⅡ was about 130 kDa. It absorbed at 223 nm. IR spectrum obtained showed that the triple helical domains of amino-acid sequences were characterized by the repetition of triplets Gly-X-Y. The MS spectrum graphically stated that CⅡextracted from cow and A. acutus have the similar peptides. The CⅡof A. acutus was obtained by extraction and purification. Appraisal analysis by SDS-PAGE, UV, IR and MS, CⅡof A. acutus was consistent with the standard CⅡ of cow. It was proved that the extracted protein was CⅡ.keywords：Agkistrodon acutus

关 键 词：蕲蛇 Ⅱ型胶原蛋白 胃蛋白酶 提取 表征 

分 类 号：R284.1[医药卫生—中药学]