title: Mitochondrial creatine kinase binding to phospholipid monolayers induces cardiolipin segregation.
abstract: It is well established that the octameric mitochondrial form of creatine kinase (mtCK) binds to the outer face of the inner mitochondrial membrane mainly via electrostatic interactions with cardiolipin (CL). However, little is known about the consequences of these interactions on membrane and protein levels. Brewster angle microscopy investigations provide, for the first time to our knowledge, images indicating that mtCK binding induced cluster formation on CL monolayers. The thickness of the clusters (10-12 nm) corresponds to the theoretical height of the mtCK-CL complex. Protein insertion into a condensed CL film, together with monolayer stabilization after protein addition, was observed by means of differential capacity measurements. Polarization modulation infrared reflection-absorption spectroscopy showed that the mean orientation of alpha-helices within the protein shifted upon CL binding from 30 degrees to 45 degrees with respect to the interface plane, demonstrating protein domain movements. A comparison of data obtained with CL and phosphatidylcholine/phosphatidylethanolamine/CL (2:1:1) monolayers indicates that mtCK is able to selectively recruit CL molecules within the mixed monolayer, consolidating and changing the morphology of the interfacial film. Therefore, CL-rich domains induced by mtCK binding could modulate mitochondrial inner membrane morphology into a raft-like organization and influence essential steps of mitochondria-mediated apoptosis.
authors.affiliation: Chemistry-Biochemistry, Université de Lyon, Lyon, France.
authors.forename: Ofelia
authors.initials: O
authors.lastname: Maniti
authors.forename: Marie-France
authors.initials: MF
authors.lastname: Lecompte
authors.forename: Olivier
authors.initials: O
authors.lastname: Marcillat
authors.forename: Bernard
authors.initials: B
authors.lastname: Desbat
authors.forename: René
authors.initials: R
authors.lastname: Buchet
authors.forename: Christian
authors.initials: C
authors.lastname: Vial
authors.forename: Thierry
authors.initials: T
authors.lastname: Granjon
mesh_terms: D000818:Animals; D002308:Cardiolipins / Q000737:chemistry / Q000378:metabolism*; D052281:Creatine Kinase, Mitochondrial Form / Q000737:chemistry / Q000378:metabolism*; D004566:Electrodes; D021962:Membrane Microdomains; D008567:Membranes, Artificial; D008853:Microscopy; D051336:Mitochondrial Membranes / Q000378:metabolism; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011487:Protein Conformation; D011817:Rabbits; D013055:Spectrophotometry, Infrared
chemical_list: D002308:Cardiolipins; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; C483858:phosphatidylethanolamine; D052281:Creatine Kinase, Mitochondrial Form