Self-assembling protein arrays on DNA chips by auto-labeling fusion proteins with a single DNA address

Proteomics. 2006 May;6(9):2650-5. doi: 10.1002/pmic.200500654.

Abstract

The high-throughput deposition of recombinant proteins on chips, beads or biosensor devices would be greatly facilitated by the implementation of self-assembly concepts. DNA-directed immobilization via conjugation of proteins to an oligonucleotide would be preeminently suited for this purpose. Here, we present a unique method to attach a single DNA address to proteins in one step during the purification from the E. coli lysate by fusion to human O6-alkylguanine-DNA-alkyltransferase (SNAP-tag) and the Avitag. Use of the conjugates in converting a DNA chip into a protein chip by self assembly is demonstrated.

MeSH terms

  • Base Sequence
  • Escherichia coli / genetics
  • Feasibility Studies
  • Molecular Probes / chemistry*
  • Molecular Sequence Data
  • Molecular Structure
  • O(6)-Methylguanine-DNA Methyltransferase / chemistry
  • Oligonucleotide Array Sequence Analysis / instrumentation
  • Oligonucleotide Array Sequence Analysis / methods*
  • Protein Array Analysis / instrumentation
  • Protein Array Analysis / methods*
  • Qb-SNARE Proteins / chemistry
  • Qc-SNARE Proteins / chemistry
  • Recombinant Fusion Proteins / chemistry*

Substances

  • Molecular Probes
  • Qb-SNARE Proteins
  • Qc-SNARE Proteins
  • Recombinant Fusion Proteins
  • SNAP23 protein, human
  • O(6)-Methylguanine-DNA Methyltransferase