Glycosylation of human pancreatic ribonuclease: differences between normal and tumor states.
Characterization of the N-glycans from human pancreatic ribonuclease (RNase 1) isolated from healthy pancreas and from pancreatic adenocarcinoma tumor cells (Capan-1 and MDAPanc-3) revealed completely different glycosylation patterns. RNase 1 from healthy cells contained neutral complex biantennary structures, with smaller amounts of tri- and tetraantennary compounds, and glycans with poly-N-acetyllactosamine extensions, all extensively fucosylated. In contrast, RNase 1 glycans from tumor cells (Capan-1) were fucosylated hybrid and complex biantennary glycans with GalNAc-GlcNAc antennae. RNase 1 glycans from Capan-1 and MDAPanc-3 cells also contained sialylated structures completely absent in the healthy pancreas. Some of these features provide distinct epitopes that were clearly detected using monoclonal antibodies against carbohydrate antigens. Thus monoclonal antibodies to Lewis(y) reacted only with normal pancreatic RNase 1, whereas, in contrast, monoclonal antibodies to sialyl-Lewis(x) and sialyl-Lewis(a) reacted only with RNase 1 secreted from the tumor cells. These glycosylation changes in a tumor-secreted protein, which reflect fundamental changes in the enzymes involved in the glycosylation pathway, open up the possibility of using serum RNase 1 as a tumor marker of pancreatic adenocarcinoma.
Peracaula
Rosa
R
Departament de Biologia, Universitat de Girona, Campus De Montilivi S/n 17071, Girona, Spain.
Royle
Louise
L
Tabares
Glòria
G
Mallorqui-Fernández
Goretti
G
Barrabés
Silvia
S
Harvey
David J
DJ
Dwek
Raymond A
RA
Rudd
Pauline M
PM
de Llorens
Rafael
R
D000230:Adenocarcinoma / Q000201:enzymology* / Q000473:pathology; D015153:Blotting, Western; D002241:Carbohydrates / Q000032:analysis; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006031:Glycosylation; D006801:Humans; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology*; D010190:Pancreatic Neoplasms / Q000201:enzymology* / Q000473:pathology; D012259:Ribonuclease, Pancreatic / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D002241:Carbohydrates; D009844:Oligosaccharides; D012259:Ribonuclease, Pancreatic