Conductometric monitoring of protein-protein interactions

J Proteome Res. 2013 Dec 6;12(12):5535-47. doi: 10.1021/pr400445v. Epub 2013 Nov 1.

Abstract

Conductometric monitoring of protein-protein and protein-sterol interactions is here proved feasible by coupling quartz crystal microbalance with dissipation monitoring (QCM_D) to nucleic acid programmable protein arrays (NAPPA). The conductance curves measured in NAPPA microarrays printed on quartz surface allowed the identification of binding events between the immobilized proteins and the query. NAPPA allows the immobilization on the quartz surface of a wide range of proteins and can be easily adapted to generate innumerous types of biosensors. Indeed multiple proteins on the same quartz crystal have been tested and envisaged proving the possibility of analyzing the same array for several distinct interactions. Two examples of NAPPA-based conductometer applications with clinical relevance are presented herein, the interaction between the transcription factors Jun and ATF2 and the interaction between Cytochrome P540scc and cholesterol.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activating Transcription Factor 2 / chemistry*
  • Activating Transcription Factor 2 / genetics
  • Activating Transcription Factor 2 / metabolism
  • Antibodies / chemistry
  • Antibodies / metabolism
  • Biosensing Techniques*
  • Cholesterol / chemistry
  • Cholesterol / metabolism
  • Cholesterol Side-Chain Cleavage Enzyme / chemistry*
  • Cholesterol Side-Chain Cleavage Enzyme / genetics
  • Cholesterol Side-Chain Cleavage Enzyme / metabolism
  • Conductometry
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Humans
  • Immobilized Proteins / chemistry
  • Immobilized Proteins / genetics
  • Immobilized Proteins / metabolism
  • Protein Array Analysis
  • Protein Binding
  • Protein Interaction Mapping
  • Proto-Oncogene Proteins c-jun / chemistry*
  • Proto-Oncogene Proteins c-jun / genetics
  • Proto-Oncogene Proteins c-jun / metabolism
  • Quartz Crystal Microbalance Techniques
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Surface Properties

Substances

  • ATF2 protein, human
  • Activating Transcription Factor 2
  • Antibodies
  • Immobilized Proteins
  • Proto-Oncogene Proteins c-jun
  • Recombinant Proteins
  • Cholesterol
  • Cholesterol Side-Chain Cleavage Enzyme