IDENTIFICATION OF CLATHRIN IN GASTRIC PARIETAL CELLS

在胃泌酸细胞中，H，K-ATP酶的运输 在根尖下管泡室和根尖之间 膜以促分泌素特异性方式发生。 底层 这个过程的机制在很大程度上是未知的。 洞察力在这 这一过程可能与植物的顶端再循环途径有关。 其他上皮细胞 我们已经通过免疫学鉴定了 泌酸细胞小管泡上的网格蛋白链。 网格蛋白是一种 已被证明在以下方面发挥作用的囊泡外壳蛋白 来自质膜的膜蛋白的内化， 在从trans-Golgi网络的运输囊泡的形成。 管囊泡上网格蛋白的鉴定是有趣的， 典型组装网格蛋白包被在 电子显微镜未观察到管状囊泡。 因此，在本发明中， 泌酸细胞管泡网格蛋白可能代表一种新的同种型 网格蛋白。 这种同种型可以以不同的方式组装， 在形态上不明显的囊状外壳中。 或者，其他 调节网格蛋白装配的外壳蛋白，如网格蛋白轻链 链和网格蛋白接头复合物，可能是独特的同种型。 因此，在本发明中， 为了帮助对所述组合物的组分进行分子表征， 含有网格蛋白的管泡状外壳，我们建议获得肽 通过质谱法从胰蛋白酶肽中获得序列。 的蛋白质 感兴趣包括：网格蛋白重链、网格蛋白轻链和 网格蛋白衔接子。 肽序列将提供直接的 目的蛋白质的表征。 另外他们可能会 用于产生抗肽抗体和寡核苷酸探针 用于目标蛋白质的分子克隆。

In gastric oxyntic cells, the trafficking of the H,K-ATPase between a subapical tubulovesicular compartment and the apical membrane occurs in a secretagogue-specific manner. The underlying mechanism of this process is largely uncharacterized. Insight in this process is likely to be relevant to the apical recycling pathway of other epithelial cells. We have immunologically identified the heavy chain of clathrin on oxyntic cell tubulovesicles. Clathrin is a vesicular coat protein that has been demonstrated to play a role in the internalization of membrane proteins from the plasma membrane and in the formation of transport vesicles from the trans-Golgi network. The identification of clathrin on tubulovesicles is intriguing, given the observation that typical assembled clathrin coats on tubulovesicles have not been observed by electron microscopy. Thus, oxyntic cell tubulovesicular clathrin may represent a novel isoform of clathrin. This isoform may assemble in a different manner resulting in a morphologically indistinct vesicular coat. Alternatively, other coat proteins modulating clathrin assembly, such as clathrin light chains and clathrin adaptor complexes, may be unique isoforms. Thus, to aid in the molecular characterization of the components of the clathrin-containing tubulovesicular coat, we propose to obtain peptide sequences from tryptic peptides by mass spectrometry. The proteins of interest include: clathrin heavy chain, clathrin light chain, and clathrin adaptors. The peptide sequences will provide immediate characterization of the proteins of interest. In addition, they may be used to generate anti-peptide antibodies and oligonucleotide probes for molecular cloning of the proteins of interest.