Demonstration of different metal ion-induced calcineurin conformations using a monoclonal antibody.
It has been suggested that calcineurin, a calmodulin-stimulated phosphatase, may exist in different metal ion-dependent conformational states (Pallen, C.J., and Wang, J. H. (1984) J. Biol. Chem. 259, 6134-6141). Evidence in favor of this hypothesis comes from studies involving a monoclonal antibody, VA1, which is specific for the small (beta) subunit of calcineurin. This antibody inhibits Ni2+-stimulated but not Mn2+-stimulated phosphatase activity against p-nitrophenyl phosphate and phosphorylase kinase. Inhibition is not due to competition of the antibody with substrate or to interference with metal ion binding to the enzyme. Complex formation between the antibody and calcineurin can be demonstrated either in the presence of Mn2+ or Ni2+ or in the absence of metal ion activators. These results indicate that the active conformational states of calcineurin are metal ion dependent, that the monoclonal antibody VA1 affects the Ni2+-induced conformational change of the enzyme, and that the beta subunit of calcineurin plays a critical role in the expression of Ni2+-stimulated phosphatase activity.
Matsui
H
H
Pallen
C J
CJ
Adachi
A M
AM
Wang
J H
JH
Lam
P H
PH
D000818:Animals; D000911:Antibodies, Monoclonal; D002147:Calmodulin / Q000494:pharmacology; D002148:Calmodulin-Binding Proteins; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D007163:Immunosorbent Techniques; D007700:Kinetics; D008345:Manganese / Q000494:pharmacology*; D009532:Nickel / Q000494:pharmacology*; D009596:Nitrophenols / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D011487:Protein Conformation / Q000187:drug effects; D051381:Rats
D000911:Antibodies, Monoclonal; D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D009596:Nitrophenols; D009943:Organophosphorus Compounds; C008644:nitrophenylphosphate; D008345:Manganese; D009532:Nickel; C022410:phosphorylase kinase phosphatase; D010749:Phosphoprotein Phosphatases