Purification of Brain-Type Creatine Kinase (B-CK) from Several Tissues of the Chicken: B-CK Subspecies

Publisher Website
Google Scholar
Abstract
A method for the purification of brain-type creatine kinase (B-CK) from several tissues of the chicken, e.g. brain, retina, gizzard and heart was developed involving (1) an affinity chromatography step on Sepharase Blue from which B-CK was specifically eluted by ADP and (2) a subsequent anion exchange chromatography step on a fast protein liquid chromatography Mono-Q column. Two distinct peaks with B-CK acivity, both purified to .gtoreq.99% homogeneity and displaying specific enzyme activities of 300-400 .mu.mol CP/min/mg lt pH 7.0 and 25.degree.C, were eluted by a salt gradient at a plateau of 150 mmol/l NaCl. The ratio of the two B-CK peaks varied in a tissue-dependent manner, indicating that in chicken the dimerization of native BB-CK from the two major B-CK subunit species is tissue-specific and nonrandom in neural tissues. The fast, efficient and convenient method for the purifcation of B-CK at small or large scale, operating at yields of 50-70%, makes the purification of this rather labile enzyme from small amounts of tissues possible and greatly facilitates the subsequent characterization of both major and minor dimeric BB-CK subspecies present in these different tissues.