Mitochondrial creatine kinase from cardiac muscle and brain are two distinct isoenzymes but both form octameric molecules.
Mitochondrial creatine kinase (Mi-CK) from chicken cardiac muscle and brain, recently shown to differ in their N-terminal amino acid sequences and to be encoded by multiple mRNAs (Hossle, H.P., Schlegel, J., Wegmann, G., Wyss, M., Böhlen, P., Eppenberger, H. M., Wallimann, T., and Perriard, J.C. (1988) Biochim. Biophys. Res. Commun. 151, 408-416) were separated on two-dimensional nonequilibrium pH-gradient electrophoresis gels and visualized as two distinct protein spots by immunoblotting. Analysis of the two proteins purified by specific elution from Blue-Sepharose with ADP (Wallimann, T., Zurbriggen, B., and Eppenberger, H. M. (1985) Enzyme 33, 226-231) followed by fast protein liquid chromatography cation exchange chromatography showed obvious differences in peptide maps, in immunological cross-reactivity with monoclonal antibodies, and in kinetic parameters. However, even though the two proteins were different, tissue-specific mitochondrial isoforms, both formed regularly-sized, perforated cube-like octameric structures with Mr of 364,000 +/- 25,000 and 352,000 +/- 20,000 for the cardiac and brain isoform, respectively. Electron microscopy of cardiac and brain Mi-CK octamers revealed cube-like molecules with a central cavity or transverse channel filled by negative stain. The octameric molecular structure of Mi-CK isoforms differs from the generally accepted dimeric arrangement of "cytosolic" muscle MM- and brain BB-CK.
Schlegel
J
J
Institute of Cell Biology, Eidgenössische Technische Hochschule Hönggerberg, Zürich, Switzerland.
Wyss
M
M
Schürch
U
U
Schnyder
T
T
Quest
A
A
Wegmann
G
G
Eppenberger
H M
HM
Wallimann
T
T
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002645:Chickens; D003402:Creatine Kinase / Q000032:analysis*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D007526:Isoelectric Point; D007527:Isoenzymes; D046911:Macromolecular Substances; D008929:Mitochondria, Heart / Q000201:enzymology*; D008970:Molecular Weight; D010449:Peptide Mapping
D007527:Isoenzymes; D046911:Macromolecular Substances; D003402:Creatine Kinase