Recovery of bands from SDS gels for MS
Sandy Kielland (kielland@UVic.CA)
Tue, 26 Jan 1999 16:39:28 -0800
Hello all;
I have a customer trying to characterize the procyclin from a Trypanosome.
She knows that she is dealing with a lot of sugar attached to a very small
peptide. There is probably a lipid anchor as well. She recently did an
arg-C digest and saw several very promising looking peaks on RP-HPLC, but
when I put them in the sequencers (ABI 473 and 475) every one of them gave
me a very large, broad peak eluting exactly where PTH-histidine elutes in
the first cycle, then nothing in subsequent cycles. My first guess was that
this might be an amino sugar, but the information I got from ABI indicates
that amino sugars will not stick to a PTH column the way they do on a PTC
column, but would zip through in the void. Has anyone out there seen
anything like this?
I have suggested that she repeat the digestion after deglycosylation, but
I'm still curious about this "bump".
cheers,
Sandy Kielland
Victoria Protein Chemistry Centre
University of Victoria
Victoria, British Columbia, Canada