title: Sensing small molecule interactions with lipid membranes by local pH modulation.
abstract: Herein, we utilized a label-free sensing platform based on pH modulation to detect the interactions between tetracaine, a positively charged small molecule used as a local anesthetic, and planar supported lipid bilayers (SLBs). The SLBs were patterned inside a flow cell, allowing for various concentrations of tetracaine to be introduced over the surface in a buffer solution. Studies with membranes containing POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) yielded an equilibrium dissociation constant value of Kd = 180 ± 47 μm for this small molecule-membrane interaction. Adding cholesterol to the SLBs decreased the affinity between tetracaine and the bilayers, while this interaction tightened when POPE (1-hexadecanoyl-2-(9-Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine) was added. Studies were also conducted with three negatively charged membrane lipids, POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (sodium salt)), POPS (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-l-serine (sodium salt)), and ganglioside GM1. All three measurements gave rise to a similar tightening of the apparent Kd value compared with pure POPC membranes. The lack of chemical specificity with the identity of the negatively charged lipid indicated that the tightening was largely electrostatic. Through a direct comparison with ITC measurements, it was found that the pH modulation sensor platform offers a facile, inexpensive, highly sensitive, and rapid method for the detection of interactions between putative drug candidates and lipid bilayers. As such, this technique may potentially be exploited as a screen for drug development and analysis.
authors.affiliation: Department of Chemistry and §Department of Biochemistry and Molecular Biology, Penn State University , University Park, PA 16802.
authors.forename: Da
authors.initials: D
authors.lastname: Huang
authors.forename: Tao
authors.initials: T
authors.lastname: Zhao
authors.forename: Wei
authors.initials: W
authors.lastname: Xu
authors.forename: Tinglu
authors.initials: T
authors.lastname: Yang
authors.forename: Paul S
authors.initials: PS
authors.lastname: Cremer
mesh_terms: D002784:Cholesterol / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000737:chemistry*; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D010713:Phosphatidylcholines / Q000737:chemistry; D013748:Tetracaine / Q000737:chemistry
chemical_list: D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D013748:Tetracaine; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine