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 mas...

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Bibliographic Details
Published in:Journal of proteome research 2004-07, Vol.3 (4), p.841-850
Main Authors: Ghosh, Dhiman, Krokhin, Oleg, Antonovici, Mihaela, Ens, Werner, Standing, Kenneth G, Beavis, Ronald C, Wilkins, John A
Format: Article
Language:English
Subjects:
Cell Membrane - chemistry
Chromatography, Affinity
Chromatography, High Pressure Liquid
Concanavalin A - chemistry
Membrane Glycoproteins - analysis
Proteomics - methods
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Wheat Germ Agglutinins - chemistry
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Summary: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 pI 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. Keywords: membrane proteins • transmembrane • MALDI MS/MS • lectin affinity chromatography • Con A • WGA • glycoproteins
ISSN:1535-3893
1535-3907
DOI:10.1021/pr049937f