Increased N-acetyl-beta-glucosaminidase activity in primary breast carcinomas corresponds to a decrease in N-acetylglucosamine containing proteins.
N-acetylglucosamine (O-GlcNAc) modification on serine or threonine residues of cytoplasmic and nuclear proteins has become a more recognized intracellular covalent modification. Removal of this modification is carried out by N-acetyl-beta-glucosaminidase (O-GlcNAcase). Since little information exists on monoglycosylation and O-GlcNAcase activity in mitogenic systems, we investigated O-GlcNAcase activity in primary breast tumors compared to matched normal adjacent breast tissue and examined enzymatic activity in relationship to the level of protein monoglycosylation. Using a variation of the acidic hexosaminidase activity assay, we demonstrated an increase in both O-GlcNAcase and lysosomal hexosaminidase activity in breast tumor tissue compared to matched adjacent tissue. Although no clear correlation with tumor grade or type was apparent among the samples examined (12 matched pairs), the increase in O-GlcNAcase and lysosomal hexosaminidase activity in tumor tissue was consistently elevated and statistically significant (P<0.05). Protein monoglycosylation was evaluated using immunoblotting, affinity blotting, and radioactive labeling. While the variety of modified proteins was greater in tumor tissue compared to adjacent tissue, the total amount of O-GlcNAc monoglycosylation was significantly decreased in the tumor tissue especially on proteins in the molecular mass range of 45-65 kDa. O-GlcNAcase may be involved in the selective removal of O-GlcNAc on certain proteins in breast tumor tissue.
Slawson
C
C
Department of Chemistry and Institute for Biomolecular Science, University of South Florida, 4202 Fowler Avenue, Tampa, FL 33620, USA.
Pidala
J
J
Potter
R
R
D000117:Acetylglucosamine / Q000032:analysis / Q000737:chemistry; D000118:Acetylglucosaminidase / Q000032:analysis / Q000378:metabolism*; D001943:Breast Neoplasms / Q000201:enzymology*; D006031:Glycosylation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D015854:Up-Regulation
D011506:Proteins; D000118:Acetylglucosaminidase; D000117:Acetylglucosamine