Deglycosylation of proteins: questions

Dale Karlson dtk at omni.cc.purdue.edu
Wed Feb 19 18:12:38 EST 1997


     My question involves the most efficient/safest method for non-
specific deglycosylation of glycoproteins.  I am not willing to
work with HF, and I have read literature concerning TFMS (Edge,
et.al 1981;  Sojar and Bahl, 1987).  Has anyone had success with
TFMS?  
     The two reviews have discrepancies concerning the usage of
anisole.  Is it necessary for the de-glycosylation?  Secondly, the
two articles also had very different "amounts" of protein necessary
for the treatment with TFMS.  Our proteins are extracted in Laemmli
buffer (or CaCl2) and are acetone precipitated. Do you know what
the minimal amount of lyophilized protein may be;  and still have
success?  Our cell-wall extraction is very time consuming (3 days; 
and in the cold room!) with a protein yield of only 280 ug/gram of
dry wood. 
     Are there any other non-specific deglycosylation methods that
you are aware of?  

                                                                                                                                                                                    Thank-you very much,
                                                                                                                                                                                    Dale Karlson



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