title: Purification and partial characterization of creatine kinase from electric organ of Electrophorus electricus (L.).
abstract: The present investigation deals with the purification and the partial characterization of the soluble creatine kinase (CK) isoenzyme, isolated from the electric organ electrocyte of Electrophorus electricus (L.). Purification was performed by precipitation of the enzyme in the crude extract with ammonium sulfate (80%). The precipitate obtained was analyzed on an ion exchange column of diethylaminoethyl cellulose-52 (DEAE) followed by gel filtration on Superose 12 in a Fast Protein Liquid Chromatography (FPLC) system. Electrophoretic mobility of the active peak confirmed previous results identifying the hybrid isoenzyme MB in the electrocyte cytoplasm. Electrocyte CK is a dimeric enzyme with two identical subunits of approximately 40 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The sequence analysis of the N-terminal peptide (14 amino acids) of the 40 kDa subunit showed homology with other CK enzymes from electric fish (Torpedo) and human muscle type CK.
authors.affiliation: Laboratório de Físico-Química Biológica, Instituto de Biofísica Carlos Chagas Filho Universidade Federal do Rio de Janeiro, RJ, Brazil. cbbsilva@biof.ufrj.br
authors.forename: C M
authors.initials: CM
authors.lastname: Batista e Silva
authors.forename: N
authors.initials: N
authors.lastname: Nunes Tavares
authors.forename: S
authors.initials: S
authors.lastname: Giovanni-De-Simone
authors.forename: A
authors.initials: A
authors.lastname: Nery da Matta
authors.forename: A
authors.initials: A
authors.lastname: Hassón-Voloch
mesh_terms: D000818:Animals; D003402:Creatine Kinase / Q000737:chemistry / Q000302:isolation & purification*; D004557:Electric Organ / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004593:Electrophorus; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D020539:Sequence Analysis, Protein
chemical_list: D007527:Isoenzymes; D003402:Creatine Kinase