| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | … k cat (min −1 ), K m (mM), k cat /K m (mM −1 min −1 ), k cat (min −1 ), K m (mM), k cat /K m (mM −1 min … CKII peptide (Ac-YPGGSTPVSSANMM-NH 2 ; 100 μM; obtained from Genscript) and UDP-GlcNAc or UDP-GlcNDAz (500 μM) were reacted with recombinant OGT (1 μM) in 20 … | Get A Quote |
The mammalian O-GlcNAc hydrolase (OGA) removes O-GlcNAc from serine and threonine residues on intracellular glycoproteins. OGA activity is sensitive to N-acyl substitutions to O-GlcNAc, with alkyl diazirine-modified O-GlcNAc (O-GlcNDAz) being completely resistant to removal by OGA. Using homology modeling, we identified OGA residues proximal to the N-acyl position of O-GlcNAc substrate. Mutation of one of these residues, C215, results in mutant enzymes that are able to hydrolytically remove O-GlcNDAz from a model compound. Further, the C215A mutant is capable of removing O-GlcNDAz from a peptide substrate. These results can be used to improve metabolism of O-GlcNAc analogs in cells. In addition, the enzyme spec... More
