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Glycoproteomics of N-glycosylation by In-Gel Deglycosylation and Matrix-Assisted Laser desorption/ionisation-time of Flight Mass Spectrometry Mapping: Application to Congenital Disorders of Glycosylation

Glycoproteomics of N-glycosylation by In-Gel Deglycosylation and Matrix-Assisted Laser desorption/ionisation-time of Flight Mass Spectrometry Mapping: Application to Congenital Disorders of Glycosylation

Dijana Sagi, Petra Kienz, Jonas Denecke, Thorsten Marquardt, Jasna Peter-Katalinić

Proteomics. 2005 Jul;5(10):2689-701.

PMID: 15912511

Abstract:

A general strategy for the structural evaluation of N-glycosylation, a common post-translational protein modification, is presented. The methods for the release of N-linked glycans from the gel-separated proteins, their isolation, purification and matrix-assisted laser desorption/ionisation-mass spectrometry (MALDI-MS) analysis of their mixtures were optimised. Since many glycoproteins are available only at low quantities from sodium dodecyl sulphate-polyacrylamide gel electrophoresis or two-dimensional gels, high attention was paid to obtain N-glycan mixtures representing their actual composition in human plasma by in-gel deglycosylation. The relative sensitivity of solid MALDI matrices for MS analysis of acidic N-glycans was compared. The most favourable results for native acidic N-glycans were obtained with 2,4,6-trihydroxyacetophenone monohydrate/diammoniumcitrate as a matrix. This matrix provided good results for both neutral and acidic mixtures as well as for methylated N-glycans. In the second part of this paper the potential of such an optimised MS strategy alone or in combination with high pH anion-exchange chromatography profiling for the clinical diagnosis of congenital disorders of glycosylation is presented.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP249278282	2′,4′,6′-Trihydroxyacetophenone monohydrate		249278-28-2	Price

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