Human brain creatine kinase binding to immobilized cibacron blue F3GA: characterization and use in purification of the enzyme.
Indian J Biochem Biophys
;
1992 Aug; 29(4): 346-9
Artículo
en Inglés
| IMSEAR
| ID: sea-27118
ABSTRACT
Creatine kinase (ATP creatine N-phosphotransferase, EC 2.7.3.2) from adult human brain grey matter was purified by cibacron blue F3GA-Sepharose affinity chromatography. By gel electrophoresis of the purified enzyme under non-denaturing conditions a single protein band was observed. The dye-bound enzyme was eluted using its substrate, ATP. Reversibility of the binding of purified creatine kinase to blue Sepharose by ATP in a concentration-dependent manner indicated that the cibacron blue molecule which structurally mimics nucleotides occupied the substrate binding site of the enzyme. Also the marked dependence of enzyme binding to blue Sepharose on Mg2+ concentration suggested that Mg2+ ion is capable of combining with the dye moiety to form a site-specific binding complex that is similar to the physiological substrate of creatine kinase, namely Mg(2+)-ATP or Mg(2+)-ADP.
Texto completo:
Disponible
Índice:
IMSEAR (Asia Sudoriental)
Asunto principal:
Triazinas
/
Encéfalo
/
Humanos
/
Bovinos
/
Cromatografía de Afinidad
/
Creatina Quinasa
/
Colorantes
/
Electroforesis en Gel de Poliacrilamida
/
Animales
/
Magnesio
Idioma:
Inglés
Revista:
Indian J Biochem Biophys
Año:
1992
Tipo del documento:
Artículo
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