Identification of N-linked carbohydrates from severe acute respiratory syndrome (SARS) spike glycoprotein.
N-glycans were released from the SARS coronavirus (SARS-CoV) spike glycoprotein produced in Vero E6 cells and their structures were determined by a combination of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry, negative ion electrospray collision-induced dissociation time-of-flight mass spectrometry and normal-phase high-performance liquid chromatography with exoglycosidase digestion. Major glycans were high-mannose (Man(5-9)GlcNAc(2)), hybrid and bi-, tri- and tetra-antennary complex with and without bisecting GlcNAc and core fucose. Complex glycans with fewer than the full complement of galactose residues were present and sialylation was negligible. Treatment with the glucosidase inhibitor N-butyl-deoxynojirimycin (NB-DNJ) inhibited N-glycan processing as evidenced by the appearance of glycans of composition Glc(3)Man(7-9)GlcNAc(2). However, some complex glycans remained suggesting the presence of an alpha-endomannosidase. Our data in tissue culture indicate that inhibition of N-glycan processing may be considered as a therapeutic strategy against SARS CoV infections.
Oxford Glycobiology Institute, Department of Biochemistry, South Parks Road, Oxford OX1 3QU, UK.
Gayle
G
Ritchie
David J
DJ
Harvey
Friederike
F
Feldmann
Ute
U
Stroeher
Heinz
H
Feldmann
Louise
L
Royle
Raymond A
RA
Dwek
Pauline M
PM
Rudd
D000818:Animals; D002241:Carbohydrates / Q000737:chemistry*; D002522:Chlorocebus aethiops; D002851:Chromatography, High Pressure Liquid; D006031:Glycosylation; D008562:Membrane Glycoproteins / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry; D045473:Severe acute respiratory syndrome-related coronavirus / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D064370:Spike Glycoprotein, Coronavirus; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry*
D002241:Carbohydrates; D008562:Membrane Glycoproteins; D011134:Polysaccharides; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV