Use of peptide microarrays for assaying phospho-dependent protein interactions

Stefano Volinia, Francesca Francioso, Lorenzo Venturoli, Lara Tosi, Mauro Marastoni, Paola Carinci, Massimo Carella, Pietro Stanziale, Rita Evangelisti

Research output: Contribution to journalArticlepeer-review

Abstract

Background. High throughput protein assays need to be devised to process and validate data produced by the current genomics projects. While DNA microarrays are now widely used in mainstream scientific experiments, protein chips are still in the developing phase. Methods. We evaluated different conditions of peptide spotting for producing peptide microarrays. Our overall aims are those of using microarray technology to assess and validate protein-protein interactions. Here we show that peptide microarrays can be used in order to assay phosphodependent and acetyl lysine dependent interactions. High affinity generic context or specific antibodies were used. Different methods of labeling and detections have been tested. Results. Optimal results were generated by using covalent linking to activated glass slides and ordinary cyanines. Conclusions. We are in the process of testing weak protein-peptide interactions by using these same peptide microarrays, and of transposing a similar technique to the study of enzyme substrates.

Original languageEnglish
Pages (from-to)281-285
Number of pages5
JournalMinerva Biotecnologica
Volume13
Issue number4
Publication statusPublished - 2001

Keywords

  • Phosphorylation
  • Post-translational modification
  • Protein array
  • Protein array analysis
  • Protein-protein interaction

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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