Bioconjugation Strategies for Connecting Proteins to DNA-Linkers for Single-Molecule Force-Based Experiments

Lyan M. van der Sleen, Katarzyna M. Tych*

*Corresponding author for this work

Research output: Contribution to journalReview articleAcademicpeer-review

2 Citations (Scopus)
17 Downloads (Pure)

Abstract

The mechanical properties of proteins can be studied with single molecule force spectroscopy (SMFS) using optical tweezers, atomic force microscopy and magnetic tweezers. It is common to utilize a flexible linker between the protein and trapped probe to exclude short-range interactions in SMFS experiments. One of the most prevalent linkers is DNA due to its well-defined properties, although attachment strategies between the DNA linker and protein or probe may vary. We will therefore provide a general overview of the currently existing non-covalent and covalent bioconjugation strategies to site-specifically conjugate DNA-linkers to the protein of interest. In the search for a standardized conjugation strategy, considerations include their mechanical properties in the context of SMFS, feasibility of site-directed labeling, labeling efficiency, and costs.

Original languageEnglish
Article number2424
Number of pages10
JournalNanomaterials
Volume11
Issue number9
DOIs
Publication statusPublished - Sep-2021

Keywords

  • optical tweezers
  • atomic force microscopy
  • single-molecule fluorescence
  • single-molecule force spectroscopy
  • unnatural amino acids
  • non-canonical amino acids
  • functionalization strategies
  • MAGNETIC TWEEZERS
  • PEPTIDE TAG
  • SPECTROSCOPY
  • TECHNOLOGY
  • BOND

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