A new functional membrane protein microarray based on tethered phospholipid bilayers

A new prototype of a membrane protein biochip is presented in this article. This biochip was created by the combination of novel technologies of peptide-tethered bilayer lipid membrane (pep-tBLM) formation and solid support micropatterning. Pep-tBLMs integrating a membrane protein were obtained in t...

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Bibliographic Details
Published in:Analyst (London) 2018-01, Vol.143 (9), p.2165-2173
Main Authors: Chadli, Meriem, Maniti, Ofelia, Marquette, Christophe, Tillier, Bruno, Cortès, Sandra, Girard-Egrot, Agnès
Format: Article
Language:English
Subjects:
Binding
Biochemistry, Molecular Biology
Chelates
Chelation
Gold
Histidine
Life Sciences
Lipid Bilayers - chemistry
Membranes
Micropatterning
Peptides
Phospholipids
Phospholipids - chemistry
Polystyrene resins
Protein Array Analysis
Proteins
Proteolipids - chemistry
Receptors, CXCR4 - chemistry
Surface Plasmon Resonance
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Summary:A new prototype of a membrane protein biochip is presented in this article. This biochip was created by the combination of novel technologies of peptide-tethered bilayer lipid membrane (pep-tBLM) formation and solid support micropatterning. Pep-tBLMs integrating a membrane protein were obtained in the form of microarrays on a gold chip. The formation of the microspots was visualized in real-time by surface plasmon resonance imaging (SPRi) and the functionality of a GPCR (CXCR4), reinserted locally into microwells, was assessed by ligand binding studies. In brief, to achieve micropatterning, P19-4H, a 4 histidine-possessing peptide spacer, was spotted inside microwells obtained on polystyrene-coated gold, and Ni-chelating proteoliposomes were injected into the reaction chamber. Proteoliposome binding to the peptide was based on metal-chelate interaction. The peptide-tethered lipid bilayer was finally obtained by addition of a fusogenic peptide (AH peptide) to promote proteoliposome fusion. The CXCR4 pep-tBLM microarray was characterized by surface plasmon resonance imaging (SPRi) throughout the building-up process. This new generation of membrane protein biochip represents a promising method of developing a screening tool for drug discovery. A new membrane protein microarray based on peptide-tethered bilayer lipid membranes formed by the fusion of cell-free expressed proteoliposomes inside micropatterned microwells.
ISSN:0003-2654
1364-5528
DOI:10.1039/c8an00260f