Stefan.Rosen at mbioekol.lu.se wrote:
: I want to enzymatically pile off the O-linked sugars (totally) in fetuin
: (from fetal calf serum). Are there anyone that have own practical
: experience about this subject and knows which enzyme(s) that works best
: (company,no, extent of deglycosylation..) ?.
: Thank you,
There is a company in Novata (o?) California, called Glyko. They have an
interesting range of recombinant glycosidases.
1. An O glycosidase that will cleave the Gal GalNAc moiety from the
Ser/Thr...HOWEVER, all substituions must be removed first, this can be
achieved using sialidases, Glyko sell one which will cleave all three
linkages (2-3, 2-6, 2-8) in combination with fucases, thus either a
cocktail approach or sequential degradation is required. Since most
O-linked sugars are much simpler in structure that N-linked, a cocktail
of 1 to 3 enzymes should do the trick. Other companies whjo might have
such enzymes are Boehringer (although they recently stopped selling
ConA..which I found a starnge marketing move?!*), and Oxford glyco
systems..on no account buy enzymes from Sigma, they are usually sourced
from elsewhere. If anyone is really desperate I can put the phone number
of GLYKO onto this newsgroup..pending thier approval, BTW I am in no way
related to GLYKO, I just like their stuff, as it is good gear.
srps at galactose.mc.duke.edu
"Insert your own pithy phrase just about here!"