Mechanism of localization of C-terminal-anchored membrane proteins

C端锚定膜蛋白的定位机制

基本信息

批准号：
09680705
负责人：
YAMAMOTO Akitsugu
金额：
$ 1.92万
依托单位：
Kansai Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680705/
关键词：
C-terminal-anchored membrane protein endoplasmic reticulum Golgi apparatus microsomal aldehyde dehydrogenase HPC-1 syntaxin 1a green fluorescent protein plasma membrane vesicular traffic GFP 局在化膜蛋白質滑面小胞体肝臓小腸選別

项目摘要

Carboxyl (C)-terminal-anchored membrane proteins are anchored to the membrane by a hydrophobic domain at its C-terminus, and play many important roles distributing on many kinds of organella such as the endoplasmic reticulum (ER), Golgi apparatus, the plasma membrane, endosomes, synaptic vesicles, mitochondria and so on. For instances, microsomal aldehyde dehydrogenase (msALDH) localizes on the ER membrane as a enzymes, while HPC-1(syntaxin 1a) localizes on the plasma membrane as an important vesicular traffic protein. In this research project, we carried out following studies in order to elucidate mechanism of the localization of the C-terminal anchored membrane proteins.1. Mechanism of the localization of HPC-1 on the plasma membrane. We investigated mechanism of localization of HPC-1 on the plasma membrane by transfecting cDNA for this protein into COS cells. As the results, it was shown that HPC-1 is integrated into ER membrane first, and then transported to the plasma membrane through Golgi apparatus. By transfecting chimeric protein with maltose binding protein (MBP) and C-terminal hydrophobic domain of HPC-1 or msALDH, it was suggested that localization signal for the plasma membrane or the ER exists on the C-terminal hydrophobic domain of HPC-1 or msALDH, respectively.2. Retention mechanism of msALDH on the ER. A Chimeric green fluorescent protein (GFP) containing the C-terminal hydrophobic domain of msALDH (GFP/msALDH) was localized on the ER when it was expressed in COS cells or NRK cells. ER localization of GFP/msALDH was not changed by AlFィイD24ィエD2ィイD1-ィエD1treatment which inhibits recycling between ER and Golgi apparatus. This suggests that GFP/msALDH is retained on the ER by static retention mechanism.3. Distribution of msALDH in rat intestine. We found that msALDH are localized on the smooth ER of the adsorptive cells of rat small intestine at high density.

羧基（C） - 末端锚定的膜蛋白被疏水结构域固定在膜上，AT是C-末端，并且在许多有机虫上扮演许多重要角色，例如内质网（例如）膜，内体，突触囊泡，例如，像卵巢一样的微粒体醛脱氢酶（MSALDH）BRANE，而HPC-1（Syntaxin 1a）位于质膜上的HPC-1（Syntaxin 1a）作为重要的囊泡蛋白质。为了进行C末端锚定膜的定位。然后，D。 HPC-1或MSALDH的末端疏水域，在EER上的保留机制。 - d24 yier d24 yier d24 yier d24 yier d24 yier d2-wie saldh被保留在静态保留机制上。在高密度下。