Dephosphorylation of tau factor by protein phosphatase 2A in synaptosomal cytosol fractions, and inhibition by aluminum.
When the synaptosomal cytosol fraction from rat brain was chromatographed on a DEAE-cellulose column and assayed for protein phosphatases for tau factor and histone H1, two peaks of activities, termed peak 1 (major) and peak 2 (minor), were separated. Each peak was in a single form 2 (minor), were separated. Each peak was in a single form on Sephacryl S-300 column chromatography. Both peaks 1 and 2 dephosphorylated tau factor phosphorylated by Ca2+/calmodulin-dependent protein kinase II and the catalytic subunit of cyclic AMP-dependent protein kinase. The Km values were in the range of 0.42-0.84 microM for tau factor. There were no differences in kinetic properties of dephosphorylation between the substrates phosphorylated by the two kinases. The phosphatase activities did not depend on Ca2+, Mn2+ and Mg2+. Immunoprecipitation and immunoblotting analysis using polyclonal antibodies to the catalytic subunit of brain protein phosphatase 2A revealed that both protein phosphatases are the holoenzymic forms of protein phosphatase 2A. Aluminum chloride inhibited the activities of both peaks 1 and 2 with IC50 values of 40-60 microM. These results suggest that dephosphorylation of tau factor in presynaptic nerve terminals is controlled mainly by protein phosphatase 2A and that the neurotoxic effect of aluminum seems to be related mostly to inhibition of dephosphorylation of tau factor.
Yamamoto
H
H
Department of Pharmacology, Kumamoto University Medical School, Japan.
Saitoh
Y
Y
Yasugawa
S
S
Miyamoto
E
E
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000535:Aluminum / Q000494:pharmacology*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D003600:Cytosol / Q000201:enzymology*; D004258:DNA Polymerase III; D011756:Diphosphates / Q000494:pharmacology; D015151:Immunoblotting; D007163:Immunosorbent Techniques; D007700:Kinetics; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D054648:Protein Phosphatase 2; D051381:Rats; D013574:Synaptosomes / Q000201:enzymology*; D014157:Transcription Factors / Q000378:metabolism*
D011756:Diphosphates; D014157:Transcription Factors; D000255:Adenosine Triphosphate; D000535:Aluminum; C041273:transcription factor tau; D004258:DNA Polymerase III; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2