Octamers of mitochondrial creatine kinase isoenzymes differ in stability and membrane binding.
Octamer stability and membrane binding of mitochondrial creatine kinase (MtCK) are important for proper functioning of the enzyme and were suggested as targets for regulatory mechanisms. A quantitative analysis of these properties, using fluorescence spectroscopy, gel filtration, and surface plasmon resonance, revealed substantial differences between the two types of MtCK isoenzymes, sarcomeric (sMtCK) and ubiquitous (uMtCK). As compared with human and chicken sMtCK, human uMtCK showed a 23-34 times slower octamer dissociation rate, a reduced reoctamerization rate and a superior octamer stability as deduced from the octamer/dimer ratios at thermodynamic equilibrium. Octamer stability of sMtCK increased with temperature up to 30 degrees C, indicating a substantial contribution of hydrophobic interactions, while it decreased in the case of uMtCK, indicating the presence of additional polar dimer/dimer interactions. These conclusions are consistent with the recently solved x-ray structure of the human uMtCK (Eder, M., Fritz-Wolf, K., Kabsch, W., Wallimann, T., and Schlattner, U. (2000) Proteins 39, 216-225). When binding to 16% cardiolipin membranes, sMtCK showed slightly faster on-rates and higher affinities than uMtCK. However, human uMtCK was able to recruit the highest number of binding sites on the vesicle surface. The observed divergence of ubiquitous and sarcomeric MtCK is discussed with respect to their molecular structures and the possible physiological implications.
Schlattner
U
U
Institute of Cell Biology, Swiss Federal Institute of Technology (ETH Zürich), Hönggerberg, CH-8093 Zürich, Switzerland. schlattn@cell.biol.ethz.ch
Wallimann
T
T
D000818:Animals; D002308:Cardiolipins / Q000378:metabolism; D002645:Chickens; D002850:Chromatography, Gel; D003402:Creatine Kinase / Q000737:chemistry* / Q000378:metabolism*; D019281:Dimerization; D004795:Enzyme Stability; D006801:Humans; D007527:Isoenzymes; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism*; D046911:Macromolecular Substances; D008928:Mitochondria / Q000201:enzymology*; D010713:Phosphatidylcholines / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012518:Sarcomeres / Q000201:enzymology; D013050:Spectrometry, Fluorescence; D020349:Surface Plasmon Resonance; D013816:Thermodynamics
D002308:Cardiolipins; D007527:Isoenzymes; D008081:Liposomes; D046911:Macromolecular Substances; D010713:Phosphatidylcholines; D011994:Recombinant Proteins; D003402:Creatine Kinase