Brain PP2A is modified by thiol-disulfide exchange and intermolecular disulfide formation.
The regulation of protein phosphatase 2A (PP2A) activity by thiol-disulfide exchange and resulting formation of an intermolecular disulfide was examined following exposure of a rat brain soluble fraction to a biotinylated derivative of the model disulfide HPDP (HPDP-biotin) which would be expected to label reactive protein thiols with a disulfide-linked biotin. The results show that a low concentration (500 microM) of HPDP-biotin produced substantial inhibition of PP2A activity and promoted the binding of the catalytic subunit of PP2A to an immobilized avidin-affinity column. Both the inhibition of PP2A activity and the binding of PP2A to the avidin column were reversed by treatment with the disulfide reducing agent dithiothreitol (DTT). Furthermore, the specific activity of PP2A was up to 7-fold higher in the DTT-eluted fractions from the avidin-affinity column than in the soluble fraction. These findings demonstrate directly that PP2A is susceptible to reversible inhibitory modification by thiol-disulfide exchange and provide mechanistic support for the emerging view that PP2A is an oxidant-sensitive protein phosphatase.
Foley
Timothy D
TD
Department of Chemistry, University of Scranton, Scranton, PA 18510, USA. foleyt2@scranton.edu
Kintner
Marissa E
ME
D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D019921:Biotinylation; D020134:Catalytic Domain; D002540:Cerebral Cortex / Q000201:enzymology / Q000378:metabolism*; D002846:Chromatography, Affinity; D004220:Disulfides / Q000378:metabolism* / Q000494:pharmacology*; D066298:In Vitro Techniques; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054648:Protein Phosphatase 2; D051381:Rats; D013438:Sulfhydryl Compounds / Q000378:metabolism* / Q000494:pharmacology*
D004220:Disulfides; C501660:N-(6-(biotinamido)hexyl)-3'-(2'-pyridyldithio)propionamide; D013438:Sulfhydryl Compounds; D001710:Biotin; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2