Interfacing lipid bilayer nanodiscs and silicon photonic sensor arrays for multiplexed protein-lipid and protein-membrane protein interaction screening.

Soluble proteins are key mediators of many biochemical signaling pathways via direct interaction with the lipid bilayer and via membrane-bound receptors. Components of the cell membrane are involved in many important biological processes, including viral infection, blood clotting, and signal transduction, and as such, they are common targets of therapeutic agents. Therefore, the development of analytical approaches to study interactions at the cell membrane is of critical importance. Herein, we integrate two key technologies, silicon photonic microring resonator arrays and phospholipid bilayer nanodiscs, which together allow multiplexed screening of soluble protein interactions with lipid and membrane-embedded targets. Microring resonator arrays are an intrinsically multiplexable, label-free analysis platform that has previously been applied to studying protein-protein, protein-nucleic acid, and nucleic acid-nucleic acid interactions. Nanodiscs are protein-stabilized lipid assemblies that represent a convenient construct to mimic the native phospholipid bilayer, investigate the effects of membrane composition, and solubilize membrane-embedded targets. Exploiting the natural affinity of nanodisc-supported lipid bilayers for oxide-passivated silicon, we assembled single and multiplex sensor arrays via direct physisorption, characterizing electrostatic effects on the nanodisc attachment. Using model systems, we demonstrate the applicability of this platform for the parallel screening of protein interactions with nanodisc-embedded lipids, glycolipids, and membrane proteins.

[1]  Abraham J. Qavi,et al.  Isothermal discrimination of single-nucleotide polymorphisms via real-time kinetic desorption and label-free detection of DNA using silicon photonic microring resonator arrays. , 2011, Analytical chemistry.

[2]  Muzammil Iqbal,et al.  Characterization of the evanescent field profile and bound mass sensitivity of a label-free silicon photonic microring resonator biosensing platform. , 2010, Biosensors & bioelectronics.

[3]  V. Gerke,et al.  Kinetics and thermodynamics of annexin A1 binding to solid-supported membranes: a QCM study. , 2002, Biochemistry.

[4]  H. Mcconnell,et al.  Allogeneic stimulation of cytotoxic T cells by supported planar membranes. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[5]  H. Pollard,et al.  Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium- and phospholipid-binding proteins. , 1994, Biochimica et biophysica acta.

[6]  S. Sligar,et al.  Monomeric Rhodopsin Is Sufficient for Normal Rhodopsin Kinase (GRK1) Phosphorylation and Arrestin-1 Binding* , 2010, The Journal of Biological Chemistry.

[7]  G. Whitesides,et al.  Noncovalent polycationic coatings for capillaries in capillary electrophoresis of proteins. , 1997, Analytical chemistry.

[8]  S. Sligar,et al.  Membrane protein assembly into Nanodiscs , 2010, FEBS letters.

[9]  Adam L. Washburn,et al.  Quantitative, label-free detection of five protein biomarkers using multiplexed arrays of silicon photonic microring resonators. , 2010, Analytical chemistry.

[10]  Stephen G. Sligar,et al.  Self-Assembly of Discoidal Phospholipid Bilayer Nanoparticles with Membrane Scaffold Proteins , 2002 .

[11]  Soo-Hyun Tark,et al.  Nanomechanical detection of cholera toxin using microcantilevers functionalized with ganglioside nanodiscs , 2010, Nanotechnology.

[12]  S. Sligar,et al.  Directed self-assembly of monodisperse phospholipid bilayer Nanodiscs with controlled size. , 2004, Journal of the American Chemical Society.

[13]  S. Sligar,et al.  Functional reconstitution of Beta2-adrenergic receptors utilizing self-assembling Nanodisc technology. , 2006, BioTechniques.

[14]  Adam L. Washburn,et al.  Sensitive on-chip detection of a protein biomarker in human serum and plasma over an extended dynamic range using silicon photonic microring resonators and sub-micron beads. , 2011, Lab on a chip.

[15]  Chang Liu,et al.  Microfluidic patterning of nanodisc lipid bilayers and multiplexed analysis of protein interaction. , 2008, Lab on a chip.

[16]  Paul Curnow,et al.  Membrane proteins, lipids and detergents: not just a soap opera. , 2004, Biochimica et biophysica acta.

[17]  J. McCann,et al.  Fluorescence analysis of galactose, lactose, and fucose interaction with the cholera toxin B subunit. , 1996, Biochemical and biophysical research communications.

[18]  M. Mrksich,et al.  Electroactive Monolayer Substrates that Selectively Release Adherent Cells , 2001, Chembiochem : a European journal of chemical biology.

[19]  Ryan C Bailey,et al.  Multiplexed evaluation of capture agent binding kinetics using arrays of silicon photonic microring resonators. , 2011, The Analyst.

[20]  S. Sligar,et al.  Chapter 11 - Reconstitution of membrane proteins in phospholipid bilayer nanodiscs. , 2009, Methods in enzymology.

[21]  Jinjun Shi,et al.  GM1 clustering inhibits cholera toxin binding in supported phospholipid membranes. , 2007, Journal of the American Chemical Society.

[22]  Ryan C Bailey,et al.  Efficient bioconjugation of protein capture agents to biosensor surfaces using aniline-catalyzed hydrazone ligation. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[23]  S. Sligar,et al.  Nanodiscs for immobilization of lipid bilayers and membrane receptors: kinetic analysis of cholera toxin binding to a glycolipid receptor. , 2008, Analytical chemistry.

[24]  J. Moss,et al.  Activation of cholera toxin by ADP-ribosylation factors, 20-kDa guanine nucleotide-binding proteins. , 1992, Current topics in cellular regulation.

[25]  Stephen G. Sligar,et al.  Defining CYP3A4 structural responses to substrate binding. Raman spectroscopic studies of a nanodisc-incorporated mammalian cytochrome P450. , 2011, Journal of the American Chemical Society.

[26]  S. Sligar,et al.  Applications of phospholipid bilayer nanodiscs in the study of membranes and membrane proteins. , 2007, Biochemistry.

[27]  Ryan C Bailey,et al.  Rapid, multiparameter profiling of cellular secretion using silicon photonic microring resonator arrays. , 2011, Journal of the American Chemical Society.

[28]  V. Gerke,et al.  Partially Reversible Adsorption of Annexin A1 on POPC/POPS Bilayers Investigated by QCM Measurements, SFM, and DMC Simulations , 2006, Chembiochem : a European journal of chemical biology.

[29]  Abraham J. Qavi,et al.  Anti-DNA:RNA antibodies and silicon photonic microring resonators: increased sensitivity for multiplexed microRNA detection. , 2011, Analytical chemistry.

[30]  Adam L. Washburn,et al.  DNA-encoding to improve performance and allow parallel evaluation of the binding characteristics of multiple antibodies in a surface-bound immunoassay format. , 2011, Analytical chemistry.

[31]  L. C. Gunn,et al.  Label-free quantitation of a cancer biomarker in complex media using silicon photonic microring resonators. , 2009, Analytical chemistry.

[32]  Matthew S. Luchansky,et al.  Silicon photonic microring resonators for quantitative cytokine detection and T-cell secretion analysis. , 2010, Analytical chemistry.

[33]  George A. Parks,et al.  The Isoelectric Points of Solid Oxides, Solid Hydroxides, and Aqueous Hydroxo Complex Systems , 1965 .

[34]  Ryan C Bailey,et al.  Nonlinear analyte concentration gradients for one-step kinetic analysis employing optical microring resonators. , 2012, Analytical chemistry.

[35]  Abraham J. Qavi,et al.  Multiplexed detection and label-free quantitation of microRNAs using arrays of silicon photonic microring resonators. , 2010, Angewandte Chemie.

[36]  E. Sackmann,et al.  Supported Membranes: Scientific and Practical Applications , 1996, Science.

[37]  W. Lencer,et al.  Membrane traffic and the cellular uptake of cholera toxin. , 1999, Biochimica et biophysica acta.

[38]  S. Sligar,et al.  Cytochromes P450 in nanodiscs. , 2011, Biochimica et biophysica acta.

[39]  Nichols Jw Phospholipid transfer between phosphatidylcholine-taurocholate mixed micelles , 1988 .

[40]  O. Zschörnig,et al.  pH and Ca2+ dependent interaction of Annexin V with phospholipid membranes: a combined study using fluorescence techniques, microelectrophoresis and infrared spectroscopy , 2000 .

[41]  Ryan C Bailey,et al.  Chaperone probes and bead-based enhancement to improve the direct detection of mRNA using silicon photonic sensor arrays. , 2012, Analytical chemistry.

[42]  D. Oprian,et al.  Transducin Activation by Nanoscale Lipid Bilayers Containing One and Two Rhodopsins* , 2007, Journal of Biological Chemistry.

[43]  James H Morrissey,et al.  The Local Phospholipid Environment Modulates the Activation of Blood Clotting* , 2007, Journal of Biological Chemistry.

[44]  Adam L. Washburn,et al.  Photonics-on-a-chip: recent advances in integrated waveguides as enabling detection elements for real-world, lab-on-a-chip biosensing applications. , 2011, The Analyst.

[45]  S. Sligar,et al.  Functional assays of membrane-bound proteins with SAMDI-TOF mass spectrometry. , 2007, Angewandte Chemie.

[46]  M. Gross,et al.  Native mass spectrometry characterization of intact nanodisc lipoprotein complexes. , 2012, Analytical chemistry.