Membrane surface-charge titration probed by gramicidin A channel conductance.
We manipulate lipid bilayer surface charge and gauge its influence on gramicidin A channel conductance by two strategies: titration of the lipid charge through bulk solution pH and dilution of a charged lipid by neutral. Using diphytanoyl phosphatidylserine (PS) bilayers with CsCl aqueous solutions, we show that the effects of lipid charge titration on channel conductance are masked 1) by conductance saturation with Cs+ ions in the neutral pH range and 2) by increased proton concentration when the bathing solution pH is less than 3. A smeared charge model permits us to separate different contributions to the channel conductance and to introduce a new method for "bilayer pKa" determination. We use the Gouy-Chapman expression for the charged surface potential to obtain equilibria of protons and cations with lipid charges. To calculate cation concentration at the channel mouth, we compare different models for the ion distribution, exact and linearized forms of the planar Poisson-Boltzmann equation, as well as the construction of a "Gibbs dividing surface" between salt bath and charged membrane. All approximations yield the intrinsic pKain of PS lipid in 0.1 M CsCl to be in the range 2.5-3.0. By diluting PS surface charge at a fixed pH with admixed neutral diphytanoyl phosphatidylcholine (PC), we obtain a conductance decrease in magnitude greater than expected from the electrostatic model. This observation is in accord with the different conductance saturation values for PS and PC lipids reported earlier (, Biochim. Biophys. Acta. 552:369-378) and verified in the present work for solvent-free membranes. In addition to electrostatic effects of surface charge, gramicidin A channel conductance is also influenced by lipid-dependent structural factors.
Rostovtseva
T K
TK
Laboratory of Physical and Structural Biology, NICHD, National Institutes of Health, Bethesda, Maryland 20892-5626 USA.
Aguilella
V M
VM
Vodyanoy
I
I
Bezrukov
S M
SM
Parsegian
V A
VA
D006096:Gramicidin / Q000737:chemistry*; D007473:Ion Channels / Q000502:physiology*; D008051:Lipid Bilayers / Q000737:chemistry*; D008564:Membrane Potentials; D008954:Models, Biological; D008958:Models, Molecular; D008962:Models, Theoretical; D008968:Molecular Conformation; D010713:Phosphatidylcholines / Q000737:chemistry; D011487:Protein Conformation; D013499:Surface Properties
D007473:Ion Channels; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D006096:Gramicidin; C021016:1,2-diphytanoylphosphatidylcholine