Rapid purification of protein phosphatase 2A from mouse brain by microcystin-affinity chromatography.
Microcystin LR, which is a monocyclic heptapeptide containing two L-amino acids, leucine and arginine, is a new inhibitor of protein phosphatases 1 and 2A. Microcystin LR-affinity chromatography was used to purify protein phosphatase 2A as a holoenzyme. Five mg of microcystin LR were immobilized to ECH Sepharose 4B by the carbodiimide coupling reaction. Following DEAE-cellulose column chromatography, microcystin-affinity chromatography, as the second step in the procedure, resulted in purification of protein phosphatase 2A in a pure form. The enzyme isolated from mouse brain consisted of two regulatory subunits of 67 kDa and 58 kDa and a catalytic subunit of 41 kDa. Microcystin-affinity chromatography is useful for isolation of protein phosphatase 2A.
Nishiwaki
S
S
Cancer Prevention Division, National Cancer Center Research Institute, Tokyo, Japan.
Fujiki
H
H
Suganuma
M
M
Nishiwaki-Matsushima
R
R
Sugimura
T
T
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D008387:Marine Toxins; D051379:Mice; D052998:Microcystins; D010456:Peptides, Cyclic; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification*; D054648:Protein Phosphatase 2
D007202:Indicators and Reagents; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; C057862:cyanoginosin LR