The relationship between glycan structures and expression levels of an endoplasmic reticulum-resident glycoprotein, UDP-glucose: Glycoprotein glucosyltransferase 1.
In this article, we report a relationship between glycan structures and expression levels of a recombinant ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1). The function of glycan structures attached to a glycoprotein is actively studied; however, the glycan structures of recombinant, and not endogenous, glycoproteins have not been examined. In this study, we indicate a relationship between the glycan structure and the level of protein expression. Expression levels were controlled utilizing a series of vectors (pFN21K, pFN22K, pFN23K, and pFN24K HaloTag CMV Flexi Vectors). Qualitative and semi-quantitative confirmation of glycan structures was achieved with tandem mass spectrometry. The results of this study indicate that glycan structures are similar to endogenous glycans at low expression levels.
ERATO, Japan Science and Technology Agency (JST), Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Shusaku
S
Daikoku
ERATO, Japan Science and Technology Agency (JST), Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Akira
A
Seko
ERATO, Japan Science and Technology Agency (JST), Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Sang-Hyun
SH
Son
ERATO, Japan Science and Technology Agency (JST), Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Katsuhiko
K
Suzuki
ERATO, Japan Science and Technology Agency (JST), Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Synthetic Cellular Chemistry Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Yukishige
Y
Ito
ERATO, Japan Science and Technology Agency (JST), Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Tokai University, Institute of Glycoscience, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan. Electronic address: kanie@tokai-u.jp.
Osamu
O
Kanie
D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D000099237:UDP-Galactose Translocators
D005964:Glucosyltransferases; D006023:Glycoproteins; D011134:Polysaccharides; D011994:Recombinant Proteins; C000731541:UGGT1 protein, human; D000099237:UDP-Galactose Translocators
Expression level; NanoLC–tandem MS; Uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase 1