Diphytanoylphosphatidylcholine dphpc

Dioleoyl-phosphatidylcholine (DOPC), dioleoyl-phosphatidylethanolamine (DOPE), diphytanoylphosphatidylcholine (DPhPC), and dierucoylphosphatidylcholine (di(C22:1)PC) were purchased from Avanti Polar Lipids, Inc. Alabaster, AL. Gramicidin A (grA) was generously gifted from O. S. Andersen, Cornell University Medical College. Bovine brain tubulin was obtained from Cytoskeleton (Denver, CO, USA). The peptide corresponding to the α-tubulin H10 domain sequence (residues 328–346), VNAAIATIKTKRSIQFVDW, was synthesized in an FDA/CBER peptide synthesis core (White Oak, MD, USA) and dissolved in DMSO. All other chemicals were analytical grade.

All the electrogenic current measurements were performed by the SURFE²R N1 (Nanion Technologies GmbH) at room temperature. All the salts were purchased from Sigma Aldrich. ATP was purchased from Life technologies. For thiolation of the chips, 1-octadecanethiol was purchased from Fluka. Electrogenic current transient measurements were performed by using solid-supported lipid membrane (SSM) technique. For forming SSM,1-Octadecanethiol was purchased from Fluka for forming monolayer over the gold surface by using Au-thiol dative bonding. Later lipid monolayer is formed from the diphytanoyl phosphatidylcholine (DPhPC) purchased from Avanti Polar lipids, USA. Next the chip surface was incubated with the sample by centrifugation at 3,000xg for 30min. Concentration jump measurements were performed by quick jump from deactivation buffer to activation buffer followed by deactivation buffer (Bazzone et al., 2013 (link); Wacker et al., 2014 (link); Barthmes et al., 2016 (link)). Transient currents due to charge transfer were measured by rapid exchange of deactivation and action buffers.