In article <420b9q$vu1 at cgl.ucsf.edu>, <neville at cgl.ucsf.edu> wrote:
> Can anybody out there point me to some good references to chemical methods of
> de-O-glycosylation of proteins or peptides. I am interested in methods
such as TFMS etc.,
> as we are trying to analyse the peptides aby MALDI.
I'm sorry if this sounds rather snotty (pointing to your own papers), but
have a look at: G.J. Rademaker, J. Haverkamp, and J. Thomas-Oates,
Determination of glycosylation sites in O-linked glycopeptides: a
sensitive mass spectrometric protocol. Org. Mass Spectrom., 1993. 28: p.
In there we describe the use of reductive beta-elimination to remove
O-glycans (and modify the amino acids to which they were attached).
Although we used FAB for analysis (and sequencing), MALDI should work OK
as well. You can compare the data you get before and after deglycosylation
(and look at the carbohydrate fraction as well)
Good luck and let me know if it works, Geert Jan
Geert Jan Rademaker
Mass Spectrometry/ Utrecht University