Bruce G, Woodley J F
Clin Chim Acta. 1981 Dec 24;117(3):325-32. doi: 10.1016/0009-8981(81)90120-0.
A simple and accurate method is described to measure the breakdown of gliadin and gliadin peptides. It involves measuring the release of the predominant amino acids glutamine and glutamic acid using a fluorometric double enzyme assay and contains none of the problems normally associated with previously used techniques. The assay is highly accurate in that small concentrations of the free amino acids can be measured with no interference from the peptide bound amino acids. The assay system was used to study the breakdown of: whole gliadin, a peptic/tryptic digest of gliadin (PT gliadin), and a peptic/tryptic/chymotryptic digest of gliadin (PTC gliadin) using a rat intestinal brush border fraction. Both PT and PTC gliadin are hydrolysed at higher rates than is whole gliadin. Exhaustive hydrolysis shows that a brush border fraction can totally break down PT gliadin while an initial linear rate of breakdown is observed (up to 60 min).
本文描述了一种简单且准确的方法来测量麦醇溶蛋白和麦醇溶蛋白肽的分解情况。该方法利用荧光双酶测定法测量主要氨基酸谷氨酰胺和谷氨酸的释放量，不存在以往技术通常会出现的问题。该测定方法非常准确，因为即使是低浓度的游离氨基酸也能被测量，且不受肽结合氨基酸的干扰。该测定系统用于研究以下物质的分解情况：完整的麦醇溶蛋白、麦醇溶蛋白的胃蛋白酶/胰蛋白酶消化物（PT麦醇溶蛋白）以及麦醇溶蛋白的胃蛋白酶/胰蛋白酶/胰凝乳蛋白酶消化物（PTC麦醇溶蛋白），使用的是大鼠肠刷状缘部分。PT和PTC麦醇溶蛋白的水解速率均高于完整麦醇溶蛋白。彻底水解表明，肠刷状缘部分能够完全分解PT麦醇溶蛋白，同时观察到最初的线性分解速率（长达60分钟）。
