In Situ Protein Microarray for Identifying the Geminivirus-Arabidopsis Interactome

Methods Mol Biol. 2024:2724:307-314. doi: 10.1007/978-1-0716-3485-1_22.

Abstract

The protein-protein interactions (PPI) by protein array technology complement other PPI assay technologies such as AP-MS and Y2H. The in situ protein array technology (NAPPA) enables low-cost, rapid, and comprehensive protein detection. It allows standardized and simultaneous assay of a wide range of proteins with a broad range of expression in cells. This technology facilitates the detection of protein-protein interactions within species and between heterologous species such as host-microbe. Here, we described the technique that identified a syntaxin-6 protein-mediated begomovirus infection using an array containing 4600 Arabidopsis genes. The protein microarray assay also identified several other viral protein-host protein interactions.

Keywords: Cell-free protein expression; HaloTag-NAPPA; Host–virus interaction; In situ protein synthesis; Protein–protein interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis*
  • Begomovirus*
  • Biological Assay
  • Geminiviridae*
  • Protein Array Analysis