Characterization and differential expression of a newly identified phosphorylated isoform of the human 20S proteasome beta7 subunit in tumor vs. normal cell lines.
The search of new pharmacological targets with original mechanism of action within the ubiquitin-proteasome pathway is still a goal to be reached in oncopharmacology. Modification by phosphorylation/dephosphorylation has been found to be involved in cancer and to regulate functional activity of proteasome. Until now, phosphorylated forms of alpha subunits of the 20S human proteasome have been mostly reported. Here, we have rationally designed a polyclonal antibody specifically directed against a phosphorylated peptide sequence bearing the beta7 subunit Ser249 residue of the human 20S proteasome. This anti-beta7 phosphoSer249 antibody appeared to be a probe of choice to detect the presence of a phosphorylated isoform of the beta7 subunit of the human 20S proteasome using mono or two-dimensional gel electrophoresis. PhosphoSer249 was sensitive to acid phosphatase treatment of native 20S proteasome. Dephosphorylation affected the peptidylglutamyl-peptide hydrolyzing activity whereas the chymotrypsin-like and trypsin-like activities remained unchanged. A comparative analysis between human normal and tumor cells showed a differential expression of the phosphoSer249 beta7 isoform with a significantly lower detection in the proteasome isolated from tumor cells, suggesting its possible use as a biomarker.
Eang
Rothmony
R
Centre de Recherche en Pharmacologie-Santé, UMR 2587 CNRS-Pierre Fabre, ISTMT, 3 rue des satellites, 31400 Toulouse, France.
Girbal-Neuhauser
Elisabeth
E
Xu
Bo
B
Gairin
Jean Edouard
JE
D015415:Biomarkers; D002460:Cell Line; D045744:Cell Line, Tumor; D002918:Chymotrypsin / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D020033:Protein Isoforms; D014357:Trypsin / Q000378:metabolism
D015415:Biomarkers; D010455:Peptides; D020033:Protein Isoforms; D002918:Chymotrypsin; D014357:Trypsin; C531344:PSMB7 protein, human; D046988:Proteasome Endopeptidase Complex