title: Kinetic control of histidine-tagged protein surface density on supported lipid bilayers.
abstract: Nickel-chelating lipids are general tools for anchoring polyhistidine-tagged proteins to supported lipid bilayers (SLBs), but controversy exists over the stability of the protein-lipid attachment. Here, we show that chelator lipids are suitable anchors for building stable, biologically active surfaces but that a simple Langmuirian model is insufficient to describe their behavior. Desorption kinetics from chelator lipids are governed by the valency of surface binding: monovalently bound proteins desorb within minutes (t1/2 approximately 6 min), whereas polyvalently bound species remain bound for hours (t1/2 approximately 12 h). Evolution between surface states is slow, so equilibrium is unlikely to be reached on experimental timescales. However, by tuning incubation conditions, the populations of each species can be kinetically controlled, providing a wide range of protein densities on SLBs with a single concentration of chelator lipid. We propose guidelines for the assembly of SLB surfaces functionalized with specific protein densities and demonstrate their utility in the formation of hybrid immunological synapses.
authors.affiliation: Department of Chemical Engineering and Department of Chemistry, University of California-Berkeley, CA 94720, USA.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Nye
authors.forename: Jay T
authors.initials: JT
authors.lastname: Groves
mesh_terms: D002614:Chelating Agents / Q000737:chemistry; D006639:Histidine / Q000737:chemistry*; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry*; D015394:Molecular Structure; D009532:Nickel / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D013499:Surface Properties; D013601:T-Lymphocytes / Q000378:metabolism
chemical_list: D002614:Chelating Agents; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D011506:Proteins; D006639:Histidine; D009532:Nickel