title: The interaction of pyrene labeled diacylglycerol with protein kinase C in mixed micelles.
abstract: The binding of protein kinase C (PKC) to pyrene-labeled diacylglycerol (pDG) has been studied in a mixed micellar system by monitoring resonance energy transfer from excited tryptophans to pyrene with time-correlated single photon counting. The average lifetime of the excited state of the tryptophans in PKC showed a clear dependence on the mole percentage pDG in micelles in contrast with pyrene-labeled phosphatidylcholine (pPC). The binding data has been analyzed to a simple model which encompasses the size of the micelles and the binding constant of the pDG-PKC complex. From our data, though, these quantities cannot be determined independently. If we have no size information on the micelles we can determine a lower boundary of this quantity compatible with the data. When the micellar size is known, a binding constant for the DG-PKC complex can be extracted. The presented analytical approach can be applied to other systems in which lipid-protein interactions must be quantified.
authors.affiliation: Department of Biochemistry, Agricultural University, Wageningen, The Netherlands.
authors.forename: P I
authors.initials: PI
authors.lastname: Bastiaens
authors.forename: E H
authors.initials: EH
authors.lastname: Pap
authors.forename: J W
authors.initials: JW
authors.lastname: Borst
authors.forename: A
authors.initials: A
authors.lastname: van Hoek
authors.forename: T
authors.initials: T
authors.lastname: Kulinski
authors.forename: R
authors.initials: R
authors.lastname: Rigler
authors.forename: A J
authors.initials: AJ
authors.lastname: Visser
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004075:Diglycerides / Q000737:chemistry / Q000378:metabolism*; D005454:Fluorescence Polarization; D007700:Kinetics; D008823:Micelles; D011493:Protein Kinase C / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D011721:Pyrenes / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D004075:Diglycerides; D008823:Micelles; D011721:Pyrenes; C030984:pyrene; D011493:Protein Kinase C