Polymer nanoarray approach for the characterization of biomolecular interactions

Sibaprasad Maity, Ekaterina Viazovkina, Alexander Gall, Yuri L. Lyubchenko

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Pair-wise interactions at the single-molecule level can be done with nanoprobing techniques, such as AFM force spectroscopy, optical tweezers, and magnetic tweezers. These techniques can be used to probe interactions between well-characterized assemblies of biomolecules, such as monomer-dimer, dimer-dimer, and trimer-monomer. An important step of these techniques is the proper assembly of dimers, trimers, and higher oligomers to enable the interactions to be probed. We have developed a novel approach in which a defined number of peptides are assembled along a flexible polymeric molecule that serves as a linear matrix, termed as flexible nanoarray (FNA). The construct is synthesized with the use of phosphoramidite chemistry (PA), in which non-nucleoside PA spacers and standard oligonucleotide synthesis are used to grow the polymeric chain with the desired length. The reactive sites are incorporated during FNA synthesis. As a result, the FNA polymer contains a set of predesigned reactive sites to which the peptides are covalently conjugated. We describe the protocol for the synthesis of FNA and the application of this methodology to measure the molecular interactions between amyloid peptides of monomer-monomer, monomer-dimer, and dimer-dimer.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages63-74
Number of pages12
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1814
ISSN (Print)1064-3745

Keywords

  • AFM
  • Amyloid oligomers
  • Click chemistry
  • Force spectroscopy
  • Nanoprobing
  • Protein-protein interactions

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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