Pauwels S, Dockray G J, Walker R, Marcus S
J Clin Invest. 1985 Jun;75(6):2006-13. doi: 10.1172/JCI111919.
The metabolism of synthetic human heptadecapeptide gastrin (G17) in vivo, and in serum in vitro, was studied by radioimmunoassay using region specific antisera, gel filtration, ion exchange chromatography, and high performance liquid chromatography. After infusion of G17 intravenously in normal human volunteers, COOH-terminal and NH2-terminal immunoreactive G17 fragments were generated. At a steady state, approximately 15% of COOH-terminal immunoreactivity was attributable to G14-like material and up to 25% of total NH2-terminal immunoreactivity was attributable to two NH2-terminal fragments; one had the chromatographic properties of 1-13 G17, and the other was less acidic and less hydrophobic. After stopping the infusion of G17, the latter fragments accounted for progressively greater proportions of total gastrin activity. When G17 was incubated in serum in vitro, there was time-dependent and temperature-dependent loss of immunoreactivity, and again COOH-terminal and NH2-terminal immunoreactive fragments were formed. Removal of the NH2-terminal pyroglutamic acid was probably the rate limiting step because synthetic 2-17 G17 was degraded more rapidly in serum (t1/2, 2-3 h) than G17 (t1/2, 3-5 h). EDTA blocked degradation at the COOH-terminus of both 2-17 G17 and G17 but cleavage at the NH2-terminus still occurred, giving rise to a G14-like peptide. The rate of conversion of G17 in serum was not enough to account for the production of fragments in vivo, and it is proposed that these are formed when G17 encounters enzymes on cell surfaces, perhaps during passage through the capillary circulation. The production of these fragments needs to be considered in interpreting studies of the identity, metabolism, and release of gastrin in health and disease.
采用区域特异性抗血清、凝胶过滤、离子交换色谱和高效液相色谱,通过放射免疫分析法研究了合成人十七肽胃泌素(G17)在体内和体外血清中的代谢情况。在正常人类志愿者静脉输注G17后,产生了COOH末端和NH2末端免疫反应性G17片段。在稳态时,约15%的COOH末端免疫反应性归因于G14样物质,高达25%的总NH2末端免疫反应性归因于两个NH2末端片段;一个具有1-13 G17的色谱特性,另一个酸性和疏水性较低。停止输注G17后,后一种片段在总胃泌素活性中所占比例逐渐增加。当G17在体外血清中孵育时,免疫反应性出现时间依赖性和温度依赖性丧失,并且再次形成了COOH末端和NH2末端免疫反应性片段。去除NH2末端焦谷氨酸可能是限速步骤,因为合成的2-17 G17在血清中的降解速度(t1/2,2-3小时)比G17(t1/2,3-5小时)更快。EDTA阻断了2-17 G17和G17在COOH末端的降解,但NH2末端的裂解仍会发生,并产生一种G14样肽。G17在血清中的转化速度不足以解释体内片段的产生,有人提出,这些片段是G17在细胞表面遇到酶时形成的,可能是在通过毛细血管循环的过程中。在解释健康和疾病状态下胃泌素的同一性、代谢和释放的研究时,需要考虑这些片段的产生情况。