Determination of the pKa of membrane-bound N,N-dimethylsphingosine using deuterium NMR spectroscopy.
Related Articles Determination of the pKa of membrane-bound N,N-dimethylsphingosine using deuterium NMR spectroscopy.
Biochim Biophys Acta. 1995 Jul 6;1237(1):37-42
Authors: Lau B, Macdonald PM
Deuterium nuclear magnetic resonance (2H-NMR) spectroscopy was applied to determine the pKa of the protein kinase C (PKC) inhibitor, N,N-dimethylsphingosine (DMS), when bound to lipid bilayers composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). The quadrupolar splittings from the deuterium labels at the alpha- and the beta-choline positions of the headgroup of POPC responded to the presence of DMS in a manner indicative of an accumulation of cationic charges near the surface plane occupied by the phospholipid phosphate group. Both quadrupolar splittings varied linearly with the amount of added DMS at pH 7.0. Conversely, at pH 10.0 DMS had virtually no influence on either quadrupole splitting, an effect attributed to titration of the dimethyl amino group of DMS to its neutral form. A DMS titration curve was obtained by quantifying the change in the quadrupolar splittings as a function of pH. The pKa of membrane-bound DMS was extracted from this 2H-NMR data by simulating the quadrupole splitting-titration curve for different values of the pKa, yielding a pKa of 8.8 after non-linear least squares fitting.
PMID: 7619840 [PubMed - indexed for MEDLINE]
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PubMed