Flip-flop of hydroxy fatty acids across the membrane as monitored by proton-sensitive microelectrodes.
Hydroxyl group-containing fatty acids play an important role in anti-inflammatory action, neuroprotection, bactericide and anti-cancer defense. However, the mechanism of long-chain hydroxy fatty acids (HFA) transport across plasma membranes is still disputed. Two main hypotheses have been suggested: firstly, that protonated HFAs traverse across the membranes spontaneously and, secondly, that the transport is facilitated by proteinaceous carriers. Here, we demonstrate that the protonated HFA are able to move across planar lipid bilayers without protein assistance. This transport step is accompanied by the acidification of the buffer in receiving compartment and the pH augmentation in the donating compartment. The latter contained liposomes doped with HFA. As revealed by scanning pH-sensitive microelectrodes, the pH shift occurred only in the immediate vicinity of the membrane, while bulk pH remained unchanged. In concurrence with the theoretical model of weak acid transport, the pH value at maximum proton flux was almost equal to the pK of the studied HFA. Intrinsic pKi values were calculated from the electrophoretic mobilities of HFA-containing liposomes and were 5.4, 6.5, 6.9 and 6.3 for 2-hydroxyhexadecanoic, 16-hydroxyhexadecanoic, 12-hydroxydodecanoic and 9,10,16-trihydroxyhexadecanoic acids, respectively.
Pohl
Elena E
EE
Institute of Cell Biology and Neurobiology, Charité-Universitätsmedizin, Charitéplatz 1, 10117 Berlin, Germany. elena.pohl@charite.de
Voltchenko
Anna M
AM
Rupprecht
Anne
A
D024202:Electrophoretic Mobility Shift Assay; D005227:Fatty Acids / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006900:Hydroxylation; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D008839:Microelectrodes; D008855:Microscopy, Electron, Scanning; D011522:Protons
D005227:Fatty Acids; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D011522:Protons