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Anal Chem. 1996 Feb 1;68(3):570-2.

Simplifying the exoglycosidase digestion/MALDI-MS procedures for sequencing N-linked carbohydrate side chains.

Author information

1
Complex Carbohydrate Research Center, University of Georgia, Athens 30602-4712, USA.

Abstract

Exoglycosidase digestion coupled with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is an effective technique for sequencing the N-linked carbohydrate side chains of a glycoprotein. However, the buffers currently used in the enzymatic procedures are detrimental to MALDI-MS, and thus desalting is required before the digestion products can be analyzed. We demonstrate that a 25 mM ammonium acetate solution adjusted to the proper pH can replace the normal exoglycosidase digestion buffers. The use of these ammonium acetate solutions permits direct MALDI-MS analysis of the digestion mixture without desalting. More importantly, we show that many of the commonly used exoglycosidases retain both their activity and their specificity under these conditions.

PMID:
8712364
DOI:
10.1021/ac950932z
[Indexed for MEDLINE]

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