Qualitative and quantitative comparison of N-glycans between pig endothelial and islet cells by high-performance liquid chromatography and mass spectrometry-based strategy.
N-glycan structures released from miniature pig endothelial and islet cells were determined by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), negative ion electrospray ionization (ESI) MS/MS and normal-phase high performance liquid chromatography (NP-HPLC) combined with exoglycosidase digestion. Totally, the identified structures were 181 N-glycans including 129 sialylated and 18 alpha-galactosylated glycans from pig endothelial cells and 80 N-glycans including 41 sialylated and one alpha-galactosylated glycans from pig islet cells. The quantity of the alpha-galactosylated glycans from pig islet cells was certainly neglectable compared to pig endothelial cells. A number of NeuGc-terminated N-glycans (80 from pig endothelial cells and 13 from pig islet cells) are newly detected by our mass spectrometric strategies. The detailed structural information will be a matter of great interest in organ or cell xenotransplantation using alpha 1,3-galactosyltransferase gene-knockout (GalT-KO) pig.
Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Korea.
Yun-Gon
YG
Kim
Geun-Cheol
GC
Gil
Kyung-Soon
KS
Jang
Sukmook
S
Lee
Hyoung-Il
HI
Kim
Jung-Sik
JS
Kim
Junho
J
Chung
Chung-Gyu
CG
Park
David J
DJ
Harvey
Byung-Gee
BG
Kim
D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D042783:Endothelial Cells / Q000737:chemistry* / Q000378:metabolism; D054794:Glycomics / Q000379:methods*; D006026:Glycoside Hydrolases / Q000378:metabolism; D007515:Islets of Langerhans / Q000737:chemistry* / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013552:Swine
D011134:Polysaccharides; D006026:Glycoside Hydrolases