Separation and identification of GM1b pathway Neu5Ac- and Neu5Gc gangliosides by on-line nanoHPLC-QToF MS and tandem MS: toward glycolipidomics screening of animal cell lines.
Monosialoganglioside fraction of YAC-1 lymphoma cells was comprehensively analyzed and structurally defined by nano-high-performance liquid chromatography (nanoHPLC) in on-line conjunction with electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-QTOF MS). An efficient separation and sensitive detection of Neu5Gc-containing gangliosides from Neu5Ac-containing analogues was for the first time accomplished in a single nanoHPLC/ESI-QTOF MS run, as demonstrated for mouse hybridoma cell GM3 fraction containing GM3(Neu5Ac) and GM3(Neu5Gc) species and further applied for the analysis of YAC-1 lymphoma cell monosialoganglioside fraction. New insights into YAC-1 monosialoganglioside mixture heterogeneity were obtained: 31 distinct species, comprising 18 Neu5Gc-containing gangliosides and 13 Neu5Ac-containing species of GM1b and GalNAc-GM1b type were found to be expressed by YAC-1 cell line. On-line structural elucidation of individually separated Neu5Ac- and Neu5Gc-containing gangliosides provided strong evidence on the "GM1b-pathway" sourcing for monosialoganglioside synthesis. Such an analytical method is documented as superior to the classical approaches by increased speed of analysis, sensitivity and level of information, being thus a viable glycolipidomic tool.
Zarei
Mostafa
M
Institute for Medical Physics and Biophysics, Robert-Koch-Str. 31, D-48149, Germany.
Müthing
Johannes
J
Peter-Katalinić
Jasna
J
Bindila
Laura
L
D000818:Animals; D002460:Cell Line; D002469:Cell Separation; D002851:Chromatography, High Pressure Liquid; D005677:G(M1) Ganglioside / Q000031:analogs & derivatives* / Q000737:chemistry; D005679:G(M3) Ganglioside / Q000737:chemistry; D006017:Glycolipids / Q000737:chemistry; D006825:Hybridomas / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D051379:Mice; D019158:N-Acetylneuraminic Acid / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D009438:Neuraminic Acids / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
D005679:G(M3) Ganglioside; D006017:Glycolipids; D008055:Lipids; D009438:Neuraminic Acids; C032592:N-glycolylneuraminic acid; D005677:G(M1) Ganglioside; C029624:ganglioside M1b; D019158:N-Acetylneuraminic Acid