N-azidoacetylmannosamine-mediated chemical tagging of gangliosides.
Peracetylated N-alpha-azidoacetylmannosamine (Ac(4)ManNAz) is metabolized by cells to CMP-azidosialic acid. It has been demonstrated previously that in this way azidosialic acid-containing glycoproteins are formed that can be labeled on the cell surface by a modified Staudinger ligation. Here, we first demonstrate that the same procedure also results in the formation of azidosialic acid-containing gangliosides. Deoxymannojirimycin, an inhibitor of N-glycan processing in proteins, decreases the total cell surface labeling in Jurkat cells by approximately 25%. Inhibition of ganglioside biosynthesis with N-[5-(adamantan-1-yl-methoxy)-pentyl]1-deoxynojirimycin reduces cell surface labeling by approximately 75%. In conclusion, exposure of cells to Ac(4)ManNAz allows in vivo chemical tagging of gangliosides.
Bussink
Anton P
AP
Department of Biochemistry, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands.
van Swieten
Paul F
PF
Ghauharali
Karen
K
Scheij
Saskia
S
van Eijk
Marco
M
Wennekes
Tom
T
van der Marel
Gijs A
GA
Boot
Rolf G
RG
Aerts
Johannes M F G
JM
Overkleeft
Herman S
HS
D017485:1-Deoxynojirimycin / Q000494:pharmacology; D001386:Azides / Q000737:chemistry / Q000378:metabolism*; D050260:Carbohydrate Metabolism / Q000187:drug effects; D005732:Gangliosides / Q000737:chemistry / Q000378:metabolism*; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D006595:Hexosamines / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism
D001386:Azides; D005732:Gangliosides; D006023:Glycoproteins; D006595:Hexosamines; C405953:N-azidoacetylmannosamine; D009844:Oligosaccharides; D017485:1-Deoxynojirimycin