Nothwehr S F, Bryant N J, Stevens T H
Division of Biological Sciences, University of Missouri, Columbia 65211, USA.
Mol Cell Biol. 1996 Jun;16(6):2700-7. doi: 10.1128/MCB.16.6.2700.
Processing of A-ALP, a late-Golgi membrane protein constructed by fusing the cytosolic domain of dipeptidyl aminopeptidase A to the transmembrane and lumenal domains of alkaline phosphatase (ALP), serves as a convenient assay for loss of retention of late-Golgi membrane proteins in Saccharomyces cerevisiae. In this study, a large group of novel grd (for Golgi retention defective) yeast mutants, representing 18 complementation groups, were identified on the basis of their mislocalization of A-ALP to the vacuole, where it was proteolytically processed and thus became enzymatically activated. All of the grd mutants exhibited significant mislocalization of A-ALP, as measured by determining the kinetics of A-ALP processing and by analyzing its
A-ALP是一种晚期高尔基体膜蛋白,通过将二肽基氨基肽酶A的胞质结构域与碱性磷酸酶(ALP)的跨膜结构域和腔结构域融合构建而成。对A-ALP的加工过程可作为一种便捷的检测方法,用于检测酿酒酵母中晚期高尔基体膜蛋白保留功能的丧失。在本研究中,基于A-ALP在液泡中的错误定位(在液泡中它被蛋白水解加工并因此被酶激活),鉴定出一大组新的grd(高尔基体保留缺陷)酵母突变体,它们代表18个互补群。通过测定A-ALP加工的动力学并分析其……,所有grd突变体均表现出A-ALP的显著错误定位。