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.nippon was extracted and purified by a combination of ammonium sulfate fractionation,ion-exchange chromatography DEAE Sepharose Fast Flow,gel filtration chromatography Sephacryl S-200,and Affi-gel Blue Gel chromatography.Results Three proteins named CNTPⅠ,CNTPⅡ,and CNTP Ⅲ were isolated.Their relative molecular weights were 6.701 8×10⁴,4.374 9×10⁴,and 2.385 5×10⁴,which were assessed by MALDI-TOF-MS and SDS-PAGE.Amino acid compositions of protein CNTPⅢ were analyzed.N-terminal Edman degradation of protein CNTPⅢ 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 CNTPⅢ is a new protein promoting cell proliferation.

国家科技部十五攻关重大项目(2004BA907A17)