Mechanisms of Peptide Transport In Human Intestinal Brush Border Membrane Vesicles

本研究的目的是利用刷状缘膜囊泡研究多肽在人小肠中的转运机制。目前，还没有关于位于人小肠上的多肽转运系统的特异性和多样性的信息。具体目的是：1.测定多肽底物的总摄取量。2.研究多肽的转运动力学。3.对多肽转运体的结构特征进行分类。这位研究人员提议进行一系列实验，首先分离从器官捐赠中获得的人类肠道刷状边界膜囊。标记底物的摄取量将通过快速过滤方法确定，多肽的水解度将使用高效液相色谱进行监测。这些多肽的简单扩散成分将通过将每个多肽视为其自身介导的转运的竞争性抑制物来确定。将进行实验以确定多肽摄取的动力学特征。为了确定转运的特异性，将使用各种二肽和三肽作为抑制剂。

The goal of this research is to study the mechanism of peptide transport in human small intestine using brush border membrane vesicles. Presently, there is no information on the specificity and multiplicity of the peptide transport systems located on human small intestine. The specific aims are: 1. To determine total uptake of the peptide substrate. 2. To investigate the kinetics of peptide transport. 3. To classify the structural characteristics of the peptide transporter. The investigator proposes to conduct a series of experiments starting with the isolation of the human intestinal brush border membrane vesicles obtained from organ donations. Uptake of labeled substrates will be determined by the rapid filtration method and hydrolysis of the peptides will be monitored using high performance liquid chromatography. The simple diffusion component of the peptides will be determined by treating each peptide as a competitive inhibitor of its own mediated transport. Experiments will be conducted to determine the kinetic characteristics of peptide uptake. Various dipeptides and tripeptides will be used as inhibitors in order to determine the specificity of transport.