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Analysis of phosphorylated isoforms of the p53 tumor suppressor protein in human lung carcinoma cells undergoing apoptosis
Author(s) -
Maxwell Steve A.,
Roth Jack A.,
Mukhopadhyay Tapas
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150171115
Subject(s) - gene isoform , apoptosis , phosphorylation , monoclonal antibody , biology , microbiology and biotechnology , flow cytometry , suppressor , antibody , biochemistry , immunology , gene
The expression of p53 tumor suppressor protein isoforms in H460a cells induced to undergo apoptosis by 2‐methoxyestradiol was investigated by two‐dimensional gel electrophoresis. Whole‐cell proteins from control and apoptotic H460a cells were separated by two‐dimensional electrophoresis and were transferred to nitrocellulose. The p53 isoforms were detected by immunoblotting using p53 monoclonal antibody Bp53‐12. Four isoforms of p53 (2, 3, 5, and 6) differing in phosphorylation state were detected in control cells. Three additional isoforms (1, 4, and 7) were observed to be expressed at significant levels only in apoptotic cells. The differential expression of isoforms 1, 4, and 7 in apoptotic cells suggests that one or more specific phosphorylation events generating these forms of p53 could play a role in regulating the function of p53 in apoptosis.