兔胃泌素的分离与鉴定

Isolation and characterization of rabbit gastrin.

作者信息

Jiang R, Huebner V D, Lee T D, Chew P, Ho F J, Shively J E, Walsh J H, Reeve J R

机构信息

First Teaching Hospital of China Medical University, Peoples Republic of China.

出版信息

Peptides. 1988 Jul-Aug;9(4):763-9. doi: 10.1016/0196-9781(88)90119-2.

DOI:10.1016/0196-9781(88)90119-2

PMID:3226952

Abstract

The heptadecapeptide form the rabbit gastrin was extracted from 16 rabbit antra and purified by a combination of DEAE Sephadex, C-18 SEP PAK cartridges, fast performance liquid chromatography (FPLC) and reverse phase high pressure liquid chromatography (HPLC) steps. After the HPLC purification, a sharp, single peak of gastrin-like immunoreactivity was detected that had the same absorption to immunoreactivity ratio as human gastrin. An amino terminal pyrrolidone carboxylic acid blocking group was removed by incubation with pyrrolidone carboxylic peptidase. The amino acid analysis, microsequence analysis and mass spectrometry all confirmed the structure of rabbit gastrin being pQGPWLQEEEEAYGWMDFamide. This sequence is identical to human gastrin-17 except for glutamine in position 6 which replaces glutamate in human gastrin. Both sulfated and unsulfated rabbit gastrin-17 were characterized by mass spectrometry.

摘要

从16只兔胃窦中提取兔胃泌素十七肽，并通过二乙氨基乙基葡聚糖（DEAE Sephadex）、C-18固相萃取柱（C-18 SEP PAK cartridges）、快速液相色谱（FPLC）和反相高压液相色谱（HPLC）步骤相结合的方法进行纯化。经过HPLC纯化后，检测到一个尖锐的、单一的胃泌素样免疫反应峰，其吸收与免疫反应的比值与人胃泌素相同。通过与焦谷氨酸肽酶孵育去除氨基末端的焦谷氨酸封闭基团。氨基酸分析、微量序列分析和质谱分析均证实兔胃泌素的结构为pQGPWLQEEEEAYGWMDFamide。该序列与人胃泌素-17相同，只是第6位的谷氨酰胺取代了人胃泌素中的谷氨酸。通过质谱对硫酸化和未硫酸化的兔胃泌素-17进行了表征。