In situ dephosphorylation of p53 protein by calf intestinal alkaline phosphatase treatment.
The monoclonal antibody (mAb) 1801 has been reported to identify an N-terminal determinant within the tumor-suppressor protein p53 located between amino acids 32 and 79. This region contains two potential sites for serine phosphorylation at amino acids 33 and 46. Using a novel technique which dephosphorylates proteins in situ in fixed permeabilized cells, we have unmasked determinants in p53 recognized by mAb 1801, allowing additional sites of p53 protein to be detected in immunohistochemically reacted cells. This result indicates that phosphorylation at one or both sites within the determinant recognized by mAb 1801 previously blocked antibody-ligand interaction. It further suggests that in situ dephosphorylation may be of more general use in identifying antibodies which can only bind to epitopes in a particular phosphorylation state.
Baunoch
D A
DA
Laboratory of Molecular Genetics, Michigan Cancer Foundation, Detroit 48201.
Xu
M
M
Tewari
A
A
Christman
J K
JK
Lane
M A
MA
D000469:Alkaline Phosphatase / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002417:Cattle; D007422:Intestines / Q000201:enzymology; D010766:Phosphorylation; D016159:Tumor Suppressor Protein p53 / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D016159:Tumor Suppressor Protein p53; D000469:Alkaline Phosphatase