TY - JOUR
T1 - Bioconjugation Strategies for Connecting Proteins to DNA-Linkers for Single-Molecule Force-Based Experiments
AU - van der Sleen, Lyan M.
AU - Tych, Katarzyna M.
N1 - Funding Information:
Funding: L.M.v.d.S. is supported by a NWO grant OCENW.Klein.526. K.M.T. is supported by a Rosalind Franklin fellowship from the University of Groningen and has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 101028366 ‘NOTE’.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/9
Y1 - 2021/9
N2 - 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.
AB - 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.
KW - optical tweezers
KW - atomic force microscopy
KW - single-molecule fluorescence
KW - single-molecule force spectroscopy
KW - unnatural amino acids
KW - non-canonical amino acids
KW - functionalization strategies
KW - MAGNETIC TWEEZERS
KW - PEPTIDE TAG
KW - SPECTROSCOPY
KW - TECHNOLOGY
KW - BOND
UR - http://www.scopus.com/inward/record.url?scp=85115084163&partnerID=8YFLogxK
U2 - 10.3390/nano11092424
DO - 10.3390/nano11092424
M3 - Review article
AN - SCOPUS:85115084163
SN - 2079-4991
VL - 11
JO - Nanomaterials
JF - Nanomaterials
IS - 9
M1 - 2424
ER -