Purification of enolase and phosphoglycerate mutase from human brain and formation of a bienzymatic complex from them |
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Authors: | KB Nazarian SZ Simonian BA Kazarian P Perez F Climent |
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Affiliation: | Barcelona University, Medical Faculty, Unit of Biochemistry, Spain. |
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Abstract: | 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. |
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