journal article·2023Open access

Evidence of Cytolysin A nanopore incorporation in mammalian cells assessed by a graphical user interface

Florian Leonardus Rudolfus Lucas, Rocio K. Finol-Urdaneta, Toon Van Thillo, Jeffrey R. McArthur, Nieck Jordy van der Heide, Giovanni Maglia, Peter Dedecker, Olaf Strauss, Carsten Wloka

Nanoscale· Volume 15· Issue 42

Abstract

Technologies capable of assessing cellular metabolites with high precision and temporal resolution are currently limited. Recent developments in the field of nanopore sensors allow the non-stochastic quantification of metabolites, where a nanopore is acting as an electrical transducer for selective substrate binding proteins (SBPs). Here we show that incorporation of the pore-forming toxin Cytolysin A (ClyA) into the plasma membrane of Chinese hamster ovary cells (CHO-K1) results in the appearance of single-channel conductance amenable to multiplexed automated patch-clamp (APC) electrophysiology. In CHO-K1 cells, SBPs modify the ionic current flowing though ClyA nanopores, thus demonstrating its potential for metabolite sensing of living cells. Moreover, we developed a graphical user interface for the analysis of the complex signals resulting from multiplexed APC recordings. This system lays the foundation to bridge the gap between recent advances in the nanopore field (e.g., proteomic and transcriptomic) and potential cellular applications.
Source: europepmc · Quality: candidate unverified · Source record ↗

Research context

OpenAlex topics: Nanopore and Nanochannel Transport Studies · Microfluidic and Capillary Electrophoresis Applications · Microfluidic and Bio-sensing Technologies

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Lab for Nanobiology, Department of Chemistry, KU Leuven, Belgium

Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia

Electrophysiology Facility for Cell Phenotyping and Drug Discovery, Wollongong, NSW 2522, Australia

Faculty of Science, Medicine and Health, University of Wollongong, Wollongong, NSW 2522, Australia

Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, 9747 AG, Groningen, The Netherlands

Experimental Ophthalmology, Department of Ophthalmology, Charité - Universitätsmedizin Berlin, A Corporate Member of Freie Universität, Humboldt-University, The Berlin Institute of Health, Berlin, Germany

Funding

Fonds Wetenschappelijk Onderzoek · Aard- en Levenswetenschappen, Nederlandse Organisatie voor Wetenschappelijk Onderzoek · Australian Research Council · Rebecca L. Cooper Medical Research Foundation

References (45)

Show source-provided references
1. Shrestha · Methods Mol. Biol. · 2020 · 2064 · 110.1007/978-1-4939-9831-9_1 ↗
2. Zenobi · Science · 2013 · 342 · 124325910.1126/science.1243259 ↗
3. Kumar · Front. Microbiol. · 2020 · 11 · 115210.3389/fmicb.2020.01152 ↗
4. Emwas · Metabolites · 2019 · 9 · 12310.3390/metabo9070123 ↗
5. Benson · Angew. Chem., Int. Ed. · 2019 · 58 · 691110.1002/anie.201900465 ↗
6. Van Thillo · Chem. Commun. · 2023 · 59 · 52010.1039/D2CC05363B ↗
7. Lucas · Angew. Chem., Int. Ed. · 2021 · 60 · 2284910.1002/anie.202107807 ↗
8. Zernia · ACS Nano · 2020 · 14 · 229610.1021/acsnano.9b09434 ↗
9. Galenkamp · ACS Catal. · 2022 · 12 · 122810.1021/acscatal.1c04388 ↗
10. Galenkamp · Nat. Chem. · 2020 · 12 · 48110.1038/s41557-020-0437-0 ↗
11. Galenkamp · Nat. Commun. · 2018 · 9 · 408510.1038/s41467-018-06534-1 ↗
12. Soskine · J. Am. Chem. Soc. · 2015 · 137 · 579310.1021/jacs.5b01520 ↗
13. Lucas · ACS Nano · 2021 · 15 · 960010.1021/acsnano.0c09958 ↗
14. Lucas · Nat. Commun. · 2021 · 12 · 579510.1038/s41467-021-26046-9 ↗
15. Huang · Nat. Commun. · 2019 · 10 · 83510.1038/s41467-019-08761-6 ↗
16. Wloka · ACS Nano · 2017 · 11 · 438710.1021/acsnano.6b07760 ↗
17. Brinkerhoff · Science · 2021 · 374 · 150910.1126/science.abl4381 ↗
18. Ouldali · Nat. Biotechnol. · 2020 · 38 · 17610.1038/s41587-019-0345-2 ↗
19. Straathof · ACS Nano · 2023 · 17 · 1368510.1021/acsnano.3c02847 ↗
20. Li · ACS Nano · 2020 · 14 · 172710.1021/acsnano.9b07385 ↗
21. Foster · Angew. Chem., Int. Ed. · 2023 · 62 · e20221456610.1002/anie.202214566 ↗
22. Ahmad · Nat. Commun. · 2023 · 14 · 137410.1038/s41467-023-36944-9 ↗
23. Cardozo · Nat. Biotechnol. · 2022 · 40 · 4210.1038/s41587-021-01002-6 ↗
24. Versloot · Nano Lett. · 2022 · 22 · 535710.1021/acs.nanolett.2c01338 ↗
25. Huang · Angew. Chem., Int. Ed. · 2022 · 61 · e20220622710.1002/anie.202206227 ↗
26. Versloot · ACS Nano · 2022 · 16 · 725810.1021/acsnano.1c11455 ↗
27. Zhang · Mol. Biomed. · 2021 · 2 · 610.1186/s43556-021-00026-3 ↗
28. Wesolowski · ChemBioChem · 2007 · 8 · 51310.1002/cbic.200600384 ↗
29. Fidzinski · Chem. Biol. · 2003 · 10 · 3510.1016/S1074-5521(02)00307-1 ↗
30. Chen · Nature · 2013 · 499 · 29510.1038/nature12354 ↗
31. Morgan · Biochem. J. · 1994 · 300 · 66510.1042/bj3000665 ↗
32. Lucas · ACS Omega · 2022 · 7 · 2604010.1021/acsomega.2c00871 ↗
33. Kobayashi · Curr. Biol. · 2018 · 28 · R38610.1016/j.cub.2018.01.007 ↗
34. Rojko · Acc. Chem. Res. · 2015 · 48 · 307310.1021/acs.accounts.5b00403 ↗
35. Sathyanarayana · Proc. Natl. Acad. Sci. U. S. A. · 2018 · 115 · E732310.1073/pnas.1721228115 ↗
36. Finol-Urdaneta · Biophys. J. · 2010 · 98 · 76210.1016/j.bpj.2009.10.053 ↗
37. Lee · Biophys. J. · 2020 · 119 · 3510.1016/j.bpj.2020.05.028 ↗
38. Houghtaling · ACS Nano · 2019 · 13 · 523110.1021/acsnano.8b09555 ↗
39. Yusko · Nat. Nanotechnol. · 2017 · 12 · 36010.1038/nnano.2016.267 ↗
40. Mutter · J. Am. Chem. Soc. · 2019 · 141 · 1435610.1021/jacs.9b06998 ↗
41. Huang · Nat. Nanotechnol. · 2015 · 10 · 98610.1038/nnano.2015.189 ↗
42. Wloka · Methods Enzymol. · 2021 · 649 · 56710.1016/bs.mie.2021.01.007 ↗
43. Bierbuesse · Nat. Commun. · 2022 · 13 · 185010.1038/s41467-022-29508-w ↗
44. Finol-Urdaneta · Proc. Natl. Acad. Sci. U. S. A. · 2022 · 11910.1073/pnas.211093211 ↗
45. Finol-Urdaneta · Mar. Drugs · 2023 · 2110.3390/md21070396 ↗