Hydroxylamine cleaveage at N/G residues of protein

"Marianne Leverone ", BIO leverone at CHUMA.CAS.USF.EDU
Wed Mar 12 11:03:44 EST 1997


Hi Newsgroup: One of the graduate students in our laboratory is trying to
cleave her protein of interest with hydroxylamine but has been unsuccesful
so far.  The protein has one site of aspargine/glycine residues.  

Protocol she has been using:
Resuspend the ppt. protein in 1% SDS Tris and incubate at 45C 30
min (to dentaure) then add 1 M hydroxylamine in 0.1 M K2CO3, pH 10,
then
incubating 45C 3 hrs.  

When she then ppts the protein and analyzes it on
SDS-PAGE, there is no change compared with untreated protein.

Has anyone out there tried hydrozylamine cleavage?  What protocol yielded
successful results?  

We would greatly appreciate anyone's help!  

Marianne, Univ. of South Florida, Tampa, Florida     




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