Single-molecule mechanical fingerprinting with DNA nanoswitch calipers.

Autor: Shrestha P; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA., Yang D; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA., Tomov TE; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA., MacDonald JI; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA., Ward A; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA., Bergal HT; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.; Biophysics Program, Harvard University, Cambridge, MA, USA., Krieg E; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA., Cabi S; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.; Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA., Luo Y; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA., Nathwani B; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA., Johnson-Buck A; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Biophysics Program, Harvard University, Cambridge, MA, USA., Shih WM; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. William_Shih@dfci.harvard.edu.; Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. William_Shih@dfci.harvard.edu.; Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA. William_Shih@dfci.harvard.edu., Wong WP; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA. Wesley.Wong@childrens.harvard.edu.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. Wesley.Wong@childrens.harvard.edu.; Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. Wesley.Wong@childrens.harvard.edu.
Jazyk: angličtina
Zdroj: Nature nanotechnology [Nat Nanotechnol] 2021 Dec; Vol. 16 (12), pp. 1362-1370. Date of Electronic Publication: 2021 Oct 21.
DOI: 10.1038/s41565-021-00979-0
Abstrakt: Decoding the identity of biomolecules from trace samples is a longstanding goal in the field of biotechnology. Advances in DNA analysis have substantially affected clinical practice and basic research, but corresponding developments for proteins face challenges due to their relative complexity and our inability to amplify them. Despite progress in methods such as mass spectrometry and mass cytometry, single-molecule protein identification remains a highly challenging objective. Towards this end, we combine DNA nanotechnology with single-molecule force spectroscopy to create a mechanically reconfigurable DNA nanoswitch caliper capable of measuring multiple coordinates on single biomolecules with atomic resolution. Using optical tweezers, we demonstrate absolute distance measurements with ångström-level precision for both DNA and peptides, and using multiplexed magnetic tweezers, we demonstrate quantification of relative abundance in mixed samples. Measuring distances between DNA-labelled residues, we perform single-molecule fingerprinting of synthetic and natural peptides, and show discrimination, within a heterogeneous population, between different posttranslational modifications. DNA nanoswitch calipers are a powerful and accessible tool for characterizing distances within nanoscale complexes that will enable new applications in fields such as single-molecule proteomics.
(© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)
Databáze: MEDLINE
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