I was using 3H-fucose for metabolic labeling of glycoproteins in chickens.
Cause I'm looking at the synaptic membrane glycoproteins, esp. the fucosylated
ones, I thought fucose would be the best precursor to use. Just to be sure, I
used double labeling with 14C-leucine, and after isolation of synaptic
membranes, I looked at the incorporation (saturation) kinetic proerties of the
two precursors. Leucine was incorporated in a text-book style fashion, leading
to saturation after ~50 min. However the fucose just did not work
(consistenltly), whatever I tried. Can anyone help me to explain these facts? Is
it possible, that I should have used 14C-fucose instead of 3H-fucose, because of
the possibility of 3H exchange with water or other molecules? Or is the fucose
"kind of labile" terminal sugar? Does anyone have experience with fucose
metabolic labeling studies? Any suggestions....
Thanx for reading this far...