RRC ID 48818
著者 Murakami-Sekimata A, Sato K, Sato K, Takashima A, Nakano A.
タイトル O-Mannosylation is required for the solubilization of heterologously expressed human beta-amyloid precursor protein in Saccharomyces cerevisiae.
ジャーナル Genes Cells
Abstract In an attempt to express human beta-amyloid precursor protein (APP) in yeast, we fortuitously found that this protein is only O-glycosylated in yeast. APP was effectively expressed in yeast, processed by yeast alpha-secretases, members of the Yapsin family, to produce N-terminal (sAPPalpha) and C-terminal (CTFalpha) domains, when its signal sequence was replaced by that of the yeast alpha-mating factor. APP is known to acquire N- and O-glycosylation through the endoplasmic reticulum (ER) and the Golgi apparatus and is transported to the plasma membrane in mammalian cells. In spite of the presence of canonical N-glycosylation consensus sequences, APP was not N-glycosylated in the yeast system. Pulse-chase experiments demonstrated that APP received only O-mannosylation in yeast. Examination of yeast pmt mutants, which are defective in the initiation of O-mannosylation in the ER, revealed that Pmt4p is most responsible for the oligosaccharide modification of APP. Maturation of APP was slowed down and aggregated forms of APP were observed by sucrose density gradient fractionation of the Deltapmt4 mutant lysate. This caused decreased production of CTFalpha. We conclude that O-mannosylation is required for the solubilization of exogenously expressed human APP.
巻・号 14(2)
ページ 205-15
公開日 2009-2-1
DOI 10.1111/j.1365-2443.2008.01263.x
PII GTC1263
PMID 19170767
MeSH Amyloid beta-Protein Precursor / chemistry* Amyloid beta-Protein Precursor / genetics Amyloid beta-Protein Precursor / metabolism* Animals COS Cells Chlorocebus aethiops Gene Expression Regulation, Fungal Glycosylation Humans Mannose / metabolism* Protein O-Methyltransferase / genetics Protein O-Methyltransferase / metabolism Protein O-Methyltransferase / physiology Protein Processing, Post-Translational / physiology* Recombinant Proteins / chemistry Recombinant Proteins / metabolism Saccharomyces cerevisiae / genetics Saccharomyces cerevisiae / metabolism* Solubility Transfection
IF 1.655
引用数 10
WOS 分野 GENETICS & HEREDITY CELL BIOLOGY
リソース情報
遺伝子材料 pTU1-PalphaAPP (RDB08714)