Glycosylation and deglycosylation of proteasomes (prosomes) from calf-liver cells: high abundance of neuraminic acid.
Proteasomes (prosomes) of calf-liver cells were probed with three different biotinylated lectins: Limulus polyphemus agglutinin (LPA), specific for neuraminic acid; Solanum tuberosum agglutinin (STA), specific for GlcNac; and concanavalin A (Con A), specific for Man/Glc. While only one proteasomal protein reacted with STA, most of the proteasomal proteins reacted with LPA and several with Con A. Deglycosylation with N-glycosidase F showed that the detected glycan residues were asparagine-linked. Finally we demonstrate an alternative method for the isolation of proteasomes based on the affinity of certain proteasomal proteins to Con A.
Schmid
H P
HP
Université Blaise Pascal, Clermont-Fd II, Aubière, France.
Vallon
R
R
Tomek
W
W
Kreutzer-Schmid
C
C
Pouch
M N
MN
Badaoui
S
S
Boissonnet
G
G
Briand
M
M
Briand
Y
Y
Buri
J
J
D000818:Animals; D060829:Arthropod Proteins; D001216:Asparagine / Q000032:analysis; D001665:Binding Sites; D005591:Chemical Fractionation; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003208:Concanavalin A / Q000494:pharmacology; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D037102:Lectins / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D009438:Neuraminic Acids / Q000032:analysis*; D037121:Plant Lectins; D011134:Polysaccharides / Q000032:analysis; D046988:Proteasome Endopeptidase Complex
D060829:Arthropod Proteins; D037102:Lectins; D009097:Multienzyme Complexes; D009438:Neuraminic Acids; D037121:Plant Lectins; D011134:Polysaccharides; C041202:potato lectin; D003208:Concanavalin A; D001216:Asparagine; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex