Multiple forms of phosphatase from human brain: isolation and partial characterization of affi-gel blue binding phosphatases.
Implication of protein phosphatases in Alzheimer disease led us to a systemic investigation of the identification of these enzyme activities in human brain. Human brain phosphatases eluted from DEAE-Sephacel with 0.22 M NaCl were resolved into two main groups by affi-gel blue chromatography, namely affi-gel blue-binding phosphatases and affi-gel blue-nonbinding phosphatases. Affi-gel blue-binding phosphatases were further separated into four different phosphatases, designated P1, P2, P3, and P4 by calmodulin-Sepharose 4B and poly-(L-lysine)-agarose chromatographies. These four phosphatases exhibited activities towards nonprotein phosphoester and two of them, P1 and P4, could dephosphorylate phosphoproteins. The activities of the four phosphatases differed in pH optimum, divalent metal ion requirements, sensitivities to various inhibitors and substrate affinities. The apparent molecular masses as estimated by gel-filtration for P1, P2, P3, and P4 were 97, 45, 42, and 125 kDa, respectively. P1 is markedly similar to PP2B from bovine brain and rabbit skeletal muscle. P4 was labeled with anti-PP2A antibody and may represent a new subtype of PP2A. P1 and P4 were also effective in dephosphorylating Alzheimer disease abnormally hyperphosphorylated tau (AD P-tau). The resulting dephosphorylated AD P-tau had its activity restored in promoting assembly of microtubules in vitro. These results suggest that P1 and P4 might be involved in the regulation of phosphorylation of tau in human brain, especially in neurodegenerative conditions like Alzheimer's disease which are characterized by the abnormal hyperphosphorylation of this protein.
Cheng
L Y
LY
Department of Neurochemistry, New York State Institute for Basic Research in Developmental Disabilities, Staten Island 10314, USA.
Wang
J Z
JZ
Gong
C X
CX
Pei
J J
JJ
Zaidi
T
T
Grundke-Iqbal
I
I
Iqbal
K
K
D000544:Alzheimer Disease / Q000201:enzymology; D001921:Brain / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D002413:Cations, Divalent; D002845:Chromatography; D004357:Drug Synergism; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D008670:Metals / Q000494:pharmacology; D008854:Microscopy, Electron; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010766:Phosphorylation; D014227:Triazines / Q000378:metabolism*; D016875:tau Proteins / Q000378:metabolism
D002147:Calmodulin; D002413:Cations, Divalent; D008670:Metals; D014227:Triazines; D016875:tau Proteins; C012842:Cibacron Blue F 3GA; D010744:Phosphoric Monoester Hydrolases; D002118:Calcium