Multicomponent Supported Membrane Microarray for Monitoring Spatially Resolved Cellular Signaling Reactions

Kabir H. Biswas, Chen Zhongwen, Alok Kumar Dubey, Dongmyung Oh, Jay T. Groves

Research output: Contribution to journalArticlepeer-review

Abstract

Cells sense biochemical as well as mechanical signals from their microenvironment by engaging multiple receptors. In many cases, multiple receptors operate in concert, and it can be misleading to attempt to isolate a single ligand–receptor interaction. Supported lipid membranes are employed to reconstitute a number of cell receptor systems. Efforts are also made to fabricate membrane microarrays presenting multiple ligands in a spatially segregated manner. However, such membrane multiplexing methods are generally limited by complex instrumentation and scalability. Here, a straightforward method is presented to produce centimeter-scale membrane microarrays displaying multiple, spatially segregated membrane-anchored protein ligands suitable for live single-cell studies. The method is based on stochastic membrane fusion and subsequent diffusion-mediated mixing of their lipid content. The result is the delivery of membrane contents into spatially segregated membrane corrals. Utilizing the technology developed here, this work probes the recruitment of an adaptor protein, Shc1, to epidermal growth factor receptor (EGFR) and EphA2 receptors and shows that activation of EGFR results in a decrease in the recruitment of protein to activate EphA2 same cell.

Original languageEnglish (US)
Article number1800015
JournalAdvanced Biosystems
Volume2
Issue number4
DOIs
StatePublished - Apr 2018
Externally publishedYes

Keywords

  • cell signaling
  • fluorescence microscopy
  • lipid exchange
  • membrane microarrays
  • microfabrication

ASJC Scopus subject areas

  • Biomaterials
  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology(all)

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