Interfacial behavior of cytoplasmic and mitochondrial creatine kinase oligomeric states.
Adsorption to the air/water interface of isoenzymes of creatine kinase was investigated using surface pressure-area isotherms and Brewster angle microscopy (BAM) observations. Octameric mitochondrial creatine kinase (mtCK) exhibits a significant affinity for the air/water interface. Whatever the mode of formation of the interfacial film, i.e., injection of the protein in the subphase or spreading onto the buffer surface, the final arrangement and conformation adopted by mtCK molecules lead to a similar result. In contrast, the dimeric isoenzymes mtCK and cytosolic MMCK do not induce any surface pressure variation. However, when the subphase contains 0.3M NaCl, both isoenzymes adsorb to the interface. When treated with 0.8 or 3M GdnHCl, muscle creatine kinase (MMCK) becomes surface active and occupies a greater surface than mtCK. This result contrasts with previous observations, often derived from monomeric proteins, that their surface activity is increased upon unfolding. It underlines the possible influence exerted by the protein oligomeric state on its interfacial activity. At a subphase pH of 8.8, which corresponds to the pI of octameric mtCK, the profiles of the isotherms obtained with dimeric and octameric states and the resistance to compression of the protein monolayers are significantly affected when compared to those recorded at pH 7.4. These data suggest that the octamer is more hydrophobic than the dimer and may contribute to explaining why octamers bind to the inner mitochondrial membrane while dimers do not.
Vernoux
Nathalie
N
UMR CNRS 5013, Biomembranes et enzymes associés, Université Claude Bernard Lyon I, 43, boulevard du 11 Novembre 1918, 69622 Villeurbanne cedex, France.
Granjon
Thierry
T
Marcillat
Olivier
O
Besson
Françoise
F
Vial
Christian
C
D000818:Animals; D003402:Creatine Kinase / Q000737:chemistry*; D052281:Creatine Kinase, Mitochondrial Form / Q000737:chemistry*; D003593:Cytoplasm / Q000737:chemistry / Q000201:enzymology*; D019281:Dimerization; D057927:Hydrophobic and Hydrophilic Interactions; D007425:Intracellular Membranes / Q000737:chemistry; D007527:Isoenzymes / Q000737:chemistry; D008853:Microscopy; D013499:Surface Properties
D007527:Isoenzymes; D003402:Creatine Kinase; D052281:Creatine Kinase, Mitochondrial Form