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Journal Articles Journal of Bioenergetics Year : 2000

A quantitative approach to membrane binding of human ubiquitous mitochondrial creatine kinase using surface plasmon resonance.

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Abstract

We have evaluated surface plasmon resonance with avidin-biotin immobilized liposomes to characterize membrane binding of ubiquitous mitochondrial creatine kinase (uMtCK). While the sarcomeric sMtCK isoform is well known to bind to negatively charged phospholipids, especially cardiolipin, this report provides the first experimental evidence on the membrane interaction of an uMtCK isoform. Qualitative measurements showed that liposomes containing 16% (w/w) cardiolipin bind octameric as well as dimeric human uMtCK and also cytochrome c, but not bovine serum albumin. Quantitative parameters could be derived only for the membrane interaction of octameric human uMtCK using an improved analytical approach. Association and dissociation kinetics of octameric uMtCK fit well to a model for heterogeneous interaction suggesting two independent binding sites. Rate constants of the two sites differed by one order of magnitude, while their affinity constants were both about 80-100 nM. The data obtained demonstrate that surface plasmon resonance with immobilized liposomes is a suitable approach to characterize the binding of peripheral proteins to a lipid bilayer and that this method yields consistent quantitative binding parameters.

Dates and versions

inserm-00390838 , version 1 (02-06-2009)

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Uwe Schlattner, Theo Wallimann. A quantitative approach to membrane binding of human ubiquitous mitochondrial creatine kinase using surface plasmon resonance.. Journal of Bioenergetics, 2000, 32 (1), pp.123-31. ⟨10.1023/A:1005576831968⟩. ⟨inserm-00390838⟩

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