The acidic environment of the Golgi is critical for glycosylation and transport.
Proteins and glycolipids are modified by various modes of glycosylation in the endoplasmic reticulum (ER) and the Golgi apparatus. It is well known that the lumen of the Golgi is acidic and compromising acidification by chemical compounds causes impaired glycosylation and transport of proteins (Axelsson et al., 2001; Chapman and Munro, 1994; Palokangas et al., 1994; Presley et al., 1997; Puri et al., 2002; Reaves and Banting, 1994; Rivinoja et al., 2006; Tartakoff et al., 1978). The mechanisms by which glycosylation and transport are regulated by an acidic pH remain largely unknown. Recent findings that the impaired regulation of an acidic environment may be implicated in the pathology of several diseases emphasize the importance of pH regulation (Jentsch, 2007; Kasper et al., 2005; Kornak et al., 2001; Kornak et al., 2008; Piwon et al., 2000; Stobrawa et al., 2001; Teichgraber et al., 2008). We recently established a mutant cell line in which Golgi acidification was selectively impaired and the raised luminal Golgi pH caused impaired transport and glycosylation of proteins and altered Golgi morphology (Maeda et al., 2008). As alkalinizing compounds nonselectively affect all acidic organelles including lysosomes, endosomes, and the Golgi, the mutant cell is thought to be useful in analyzing how the acidic environment of the Golgi regulates glycosylation. In this chapter, we have introduced how we established mutant cells with impaired Golgi acidification and methods for measuring Golgi pH.
Maeda
Yusuke
Y
Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan.
Kinoshita
Taroh
T
D000143:Acids / Q000494:pharmacology*; D000818:Animals; D018929:Cell Culture Techniques; D017930:Genes, Reporter; D005821:Genetic Techniques; D006031:Glycosylation / Q000187:drug effects; D006056:Golgi Apparatus / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008954:Models, Biological; D016296:Mutagenesis / Q000502:physiology; D011499:Protein Processing, Post-Translational / Q000187:drug effects / Q000235:genetics; D021381:Protein Transport / Q000187:drug effects / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods
D000143:Acids; D011506:Proteins