The effect of phosphorylation on the antigenic reactivity of p53 in cultured human keratinocytes.
The detection of p53 in human keratinocytes is dependent on the specific anti-p53 monoclonal antibody that is used. Differences in antibody recognition are postulated to be due to the masking or exposure of particular epitopes in different conformations of p53. This study addresses the role of phosphorylation on p53-epitope accessibility in human keratinocytes. Keratinocytes were treated with the phosphatase inhibitor, okadaic acid, to determine the effect of inhibiting cellular phosphatases on p53 phosphorylation and epitope recognition. These studies suggest there is a correlation between the level of p53 phosphorylation and the antigenic reactivity of certain p53 epitopes in human keratinocytes. We also examined the ability of the catalytic subunits of protein phosphatase 1 and 2A to dephosphorylate p53 derived from human keratinocytes in vitro. These data suggest that PP2A may be the phosphatase that acts on p53 in cultured human keratinocytes.
Kumar
M
M
Department of Dermatology, Indiana University School of Medicine, Indianapolis 46202, USA.
Spandau
D F
DF
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002478:Cells, Cultured; D000939:Epitopes / Q000032:analysis*; D004988:Ethers, Cyclic / Q000494:pharmacology; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000378:metabolism*; D046911:Macromolecular Substances; D008297:Male; D019319:Okadaic Acid; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation; D054645:Protein Phosphatase 1; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000032:analysis* / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D004988:Ethers, Cyclic; D046911:Macromolecular Substances; D010750:Phosphoproteins; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53; D019319:Okadaic Acid; D054645:Protein Phosphatase 1; D017027:Protein Tyrosine Phosphatases