The catalytic subunit of protein phosphatase 2A is carboxyl-methylated in vivo.
We have used polyclonal antibodies against an internal peptide (residues 169 to 182; Ab169/182) and a peptide corresponding to the carboxyl terminus (residues 299 to 309; Ab299/309) to look for in vivo modifications of protein phosphatase 2A catalytic (PP2Ac) subunit. Treatment of extracts from human breast cancer (MCF7) cells with either alkali or ethanol increased immunoreactivity of PP2Ac subunit severalfold on Western blots with Ab299/309, but did not apparently change molecular weight or isoelectric point of the protein. In contrast, immunoreactivity with Ab169/182 was unchanged by these treatments. Subsequently, we demonstrated that the increase in PP2Ac subunit recognition by Ab299/309 coincides with the demethylation of this protein at the carboxyl-terminal leucine (Leu309). Methylation of PP2Ac subunit, in vitro, increases its activity toward both phosphorylase a and a phosphopeptide. The carboxyl-terminal sequence (TPDYFL) of PP2Ac subunit is completely conserved between mammals, yeast, fruit fly, and plants which suggests that regulation of this enzyme activity by carboxyl-terminal methylation has been conserved during evolution.
Favre
B
B
Friedrich Miescher-Institut, Basel, Switzerland.
Zolnierowicz
S
S
Turowski
P
P
Hemmings
B A
BA
D000595:Amino Acid Sequence; D001943:Breast Neoplasms; D002384:Catalysis; D017124:Conserved Sequence; D005260:Female; D006801:Humans; D007930:Leucine / Q000031:analogs & derivatives; D008745:Methylation; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D010762:Phosphorylase a / Q000378:metabolism; D010766:Phosphorylation; D011496:Protein Methyltransferases / Q000378:metabolism*; D054648:Protein Phosphatase 2; D011499:Protein Processing, Post-Translational; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D009842:Oligopeptides; C037642:kemptide; D011496:Protein Methyltransferases; D010762:Phosphorylase a; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D007930:Leucine