梅花鹿茸中活性多肽的纯化、测序及功能研究

Purification, Sequencing and Biological Activity of Polypeptide from Velvet Antler

利用凝胶过滤、离子交换层析及C18反相柱层析等方法,从梅花鹿茸中分离得到了一个多肽CNT14,SDS-PAGE电泳显示其为一条带,HPLC图谱为单峰,激光解析电离飞行时间质谱测定其分子量为1479.9028.氨基酸组成分析结果表明,此多肽主要含缬氨酸、亮氨酸、谷氨酸及脯氨酸,用电喷雾串联质谱法测序结果为E-P-T-V-L-D-E-V-C-L-A-H-G-P.活性检测结果表明,该多肽能够明显促进小鼠海马HT22细胞增殖.同时,采用固相合成法对该多肽进行了合成,与所分离的天然多肽进行了结构与性质的比较.

The velvet antler polypeptide CNT14 was extracted and purified by gel filtration, ion exchange chromatography and RP C18, which showed a single peak in HPLC chromatography and a single band in SDSPAGE. The molecular weight measured by MALDI/TOF/MS spectrum was 1479. 9028. The polypeptide consisted mostly of Glu, Leu, Val, Pro. The amino acid sequence of the polypeptide was detected with ESI-MS/ MS, and the sequence was E-P-T-V-L-D-E-V-C-L-A-H-G-P. The experiments of biological activity of poly- peptide CNT14 in vivo were carried out, and the results show that CNT14 has stimulant effects on the growth of rat HT22 cells. Then we produced the polypeptide CNT14 according the amino acid sequence by solid phase synthesis, confirmed the sequence of the polypeptide to be consistent with the amino acid sequence of polypeptide CNT14 which was separated from the velvet antler.