Cantilever sensor for nanomechanical detection of specific protein conformations

Nano Lett. 2005 Dec;5(12):2385-8. doi: 10.1021/nl051449z.

Abstract

A fast, label-free, and multiplexed method based on piezoresistive cantilevers is reported for the detection of specific protein conformations at the nanoscale level. The ligand-binding domain of the human oestrogen receptor (ERalpha-LBD) is used as the experimental model system, and ERalpha-LBD with or without oestradiol (E2) is detected using the conformation-specific peptides alpha/betaI (Ser-Ser-Asn-His-Gln-Ser-Ser-Arg-Leu-Ile-Glu-Leu-Leu-Ser-Arg, which recognizes E2-bound ER) and alpha/betaII (Ser-Ala-Pro-Arg-Ala-Thr-Ile-Ser-His-Tyr-Leu-Met-Gly-Gly, which recognizes E2-free ER). Target-specific signals are obtained in situ at protein concentrations of 2.5-20 nM. The in-build electrical readout of the piezoresistive cantilevers provides a convenient alternative to the conventional optical detection, and the presented method offers the possibility of detecting protein conformational changes using miniaturized microarrays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / instrumentation*
  • Biosensing Techniques / methods
  • Electrochemistry / instrumentation*
  • Electrochemistry / methods
  • Equipment Design
  • Equipment Failure Analysis
  • Estradiol / chemistry*
  • Estrogen Receptor alpha / analysis
  • Estrogen Receptor alpha / chemistry*
  • Mechanics
  • Microchemistry / instrumentation
  • Microchemistry / methods
  • Nanotechnology / instrumentation*
  • Nanotechnology / methods
  • Protein Array Analysis / instrumentation*
  • Protein Array Analysis / methods
  • Protein Binding
  • Protein Conformation
  • Protein Interaction Mapping / instrumentation*
  • Protein Interaction Mapping / methods
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Estrogen Receptor alpha
  • Estradiol