梅花鹿鹿茸中促进海马神经细胞增殖蛋白的分离纯化  

Separation and purification of protein promoting hippocampus nerve cell proliferation from pilose antler of Cervus nippon

作　　者：严铭铭[1] 曲晓波[1] 钟英杰[1] 赵大庆[1] 刘宁[2] 刘志强[2] 刘淑莹[2] 

机构地区：[1]长春中医药大学研发中心,吉林长春130117 [2]中国科学院长春应用化学研究所,吉林长春130023

出　　处：《中草药》2007年第8期1163-1167,共5页Chinese Traditional and Herbal Drugs

基　　金：国家科技部十五攻关重大项目(2004BA907A17)

摘　　要：目的研究梅花鹿鹿茸中蛋白的分离纯化和促进细胞增殖的生物活性。方法采用硫酸铵分级沉淀法提取鹿茸蛋白,蛋白组分经过DEAE Sep Harose FastFlow、SepHacrylS-200和Affi-gel Blue Gel柱色谱分离纯化。结果分离得到3个单一蛋白化合物CNTP Ⅰ、CNTP Ⅱ、CNTP Ⅲ。结合激光解析电离飞行时间质谱和SDS-聚丙烯酰胺凝胶电泳确定其相对分子质量分别为6.7018×104、4.3749×104、2.3855×104。分析了CNTPⅢ的氨基酸组成,采用Edman测序法测定N末端氨基酸序列为GDRGTAAKHALDEEP。CNTP Ⅲ具有促进小鼠海马神经细胞(HT22)增殖的作用。结论 CNTPⅢ是具有促进细胞增殖的新蛋白。Objective To investigate the separation and purification of the proteins from pilose antler of Cervus nippon and their cell proliferation promoting bioactivities. Methods The pilose antler of C. nip- pon was extracted and purified by a combination of ammonium sulfate fractionation, ion-exchange chro- matography DEAE Sepharose Fast Flow, gel filtration chromatography Sephacryl S-200, and Affi-gel Blue Gel chromatography. Results Three proteins named CNTP I , CNTPⅡ , and CNTP Ⅲ were isolated. Their relative molecular weights were 6.7018×10^4, 4.374 9×10^4, and 2.3855×10^4, which were assessed by MALDI-TOF-MS and SDS-PAGE. Amino acid compositions of protein CNTP Ⅲ were ana-lyzed. N-terminal Edman degradation of protein CNTP Ⅲ showed its amino acid sequence GDRGTAAKHALDEEP. The bioactivity tests of protein CNTP Ⅲ showed significantly stimulant effects on the proliferation of nerve cells HT22 in mice. Conclusion Protein CNTP Ⅲ is a new protein promoting cell proliferation.

关 键 词：梅花鹿 鹿茸 蛋白 生物活性 

分 类 号：R286.1[医药卫生—中药学]