中华绒螯蟹促性腺释放激素类似物的初步分离纯化与免疫鉴定

摘要

应用反向高效液相色谱法(RP-HPLC)从中华绒螯蟹脑组织提取液初步分离了促性腺释放激素(GnRH)类似物,使用章鱼促性腺释放激素抗体(anti-octGnRH)对分离组分进行免疫斑点印迹实验鉴定,获得了免疫反应呈阳性分离组分,表明脑组织粗提液中含有GnRH 类似物分离组分.对该GnRH类似物进行基质辅助激光解吸/电离串联飞行时间质谱(MALDI-TOF/TOF MS)分析,结果显示多肽物质分子量范围在800～1400 u,主要由8种小分子多肽组成,其中一种组分的分子量与章鱼促性腺释放激素(octGnRH)分子量接近.%In this study a gonadotropin-releasing hormone (GnRH)-like peptide was partially isolated by reverse phase high performance liquid chromatography (RP-HPLC) from the brain extract of Chinese mitten crab Eriocheir sinensis. Immunological dot blot analyses from RP-HPLC fractions using the an-ti-octGnRH revealed that one of fractions is immune reaction positive, suggesting that the fractions of brain extract contain GnRH-like substance. Eight kinds of small peptides, with molecular weight ranging from 800 to 1 400 u, were further identified in this fraction by using matrix-assisted laser desorp-tion/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF MS). One of peptides in MS component shared similar molecular weight with the octGnRH.