Lectin affinity as an approach to the proteomic analysis of membrane glycoproteins.
The aim was to determine the proportion of membrane glycoproteins captured using concanavalin A or wheat germ agglutinin lectin affinity chromatography. Digests of the isolated proteins were separated by reversed-phase liquid chromatography and analyzed by matrix-assisted laser desorption tandem mass spectrometry. The two lectins identified different groups of proteins with a broad range of molecular mass and p/ values, including a number of proteins that overlapped the two groups. Approximately 30% of the proteins were positively identified as containing domains that were predicted using standard bioinformatics methods to be characteristic of integral membrane proteins. This approach represents an effective method of surveying the membrane protein pool of mammalian cells for subsequent proteomic analysis.
Ghosh
Dhiman
D
Manitoba Centre for Proteomics, Rheumatic Disease Research Laboratory, University of Manitoba, Canada.
Krokhin
Oleg
O
Antonovici
Mihaela
M
Ens
Werner
W
Standing
Kenneth G
KG
Beavis
Ronald C
RC
Wilkins
John A
JA
D002462:Cell Membrane / Q000737:chemistry; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003208:Concanavalin A / Q000737:chemistry*; D008562:Membrane Glycoproteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014909:Wheat Germ Agglutinins / Q000737:chemistry*
D008562:Membrane Glycoproteins; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A