[Purification of enolase and phosphoglycerate mutase from human brain and formation of a bienzymatic complex from them].
Neuron-specific enolase and phosphoglycerate mutase with specific activities of 106 and 215 U/mg, respectively, have been purified from human brain. Hydrophobic chromatography for enolase and blue Sepharose affinity chromatography for phosphoglycerate mutase were used as the last steps of purification. A heterobifunctional complex with fully preserved enolase and phosphoglycerate mutase activities was synthesized with the use of a bifunctional reagent, N-succinimidyl 3-(2-pyridyldithio)propionate. Autoantibodies to the conjugate will be used for identifying the bienzymatic complex in vivo.
Nazarian
K B
KB
Barcelona University, Medical Faculty, Unit of Biochemistry, Spain.
Simonian
S Z
SZ
Kazarian
B A
BA
Perez
P
P
Climent
F
F
D001323:Autoantibodies / Q000378:metabolism; D001921:Brain / Q000201:enzymology*; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents; D006801:Humans; D009097:Multienzyme Complexes / Q000276:immunology / Q000378:metabolism*; D010736:Phosphoglycerate Mutase / Q000302:isolation & purification* / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000302:isolation & purification* / Q000378:metabolism; D013388:Succinimides
D001323:Autoantibodies; D003432:Cross-Linking Reagents; D009097:Multienzyme Complexes; D013388:Succinimides; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; D010751:Phosphopyruvate Hydratase; D010736:Phosphoglycerate Mutase
Ochistka enolazy i fosfoglitseratmutazy iz mozga cheloveka i formirovanie iz nik bifermentnogo kompleksa.