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Bertheussen, K.; Plassche, M.A.T. van de; Bakkum, T.; Gagestein, B.; Ttofi, I.; Sarris, A.J.C.; ... ; Kasteren, S.I. van (2022)

Live-cell imaging of sterculic acid - a naturally occurring 1,2-cyclopropene fatty acid - by bioorthogonal reaction with turn-on tetrazine-fluorophore conjugates

Article / Letter to editor
In the field of lipid research, bioorthogonal chemistry has made the study of lipid uptake and processing in living systems possible, whilst minimising biological properties arising from detectable pendant groups. To allow the study of unsaturated free fatty acids in live cells, we here report the use of sterculic acid, a 1,2-cyclopropene-containing oleic acid analogue, as a bioorthogonal probe. We show that this lipid can be readily taken up by dendritic cells without toxic side effects, and that it can subsequently be visualised using an inverse electron-demand Diels-Alder reaction with quenched tetrazine-fluorophore conjugates. In addition, the lipid can be used to identify changes in protein oleoylation after immune cell activation. Finally, this reaction can be integrated into a multiplexed bioorthogonal reaction workflow by combining it with two sequential copper-catalysed Huisgen ligation reactions. This allows for the study of multiple biomolecules in the cell simultaneously...Show moreIn the field of lipid research, bioorthogonal chemistry has made the study of lipid uptake and processing in living systems possible, whilst minimising biological properties arising from detectable pendant groups. To allow the study of unsaturated free fatty acids in live cells, we here report the use of sterculic acid, a 1,2-cyclopropene-containing oleic acid analogue, as a bioorthogonal probe. We show that this lipid can be readily taken up by dendritic cells without toxic side effects, and that it can subsequently be visualised using an inverse electron-demand Diels-Alder reaction with quenched tetrazine-fluorophore conjugates. In addition, the lipid can be used to identify changes in protein oleoylation after immune cell activation. Finally, this reaction can be integrated into a multiplexed bioorthogonal reaction workflow by combining it with two sequential copper-catalysed Huisgen ligation reactions. This allows for the study of multiple biomolecules in the cell simultaneously by multimodal confocal imaging.Show less
All authors
Bertheussen, K.; Plassche, M.A.T. van de; Bakkum, T.; Gagestein, B.; Ttofi, I.; Sarris, A.J.C.; Overkleeft, H.S.; Stelt, M. van der; Kasteren, S.I. van
Date
2022-07-15
Journal
Angewandte Chemie (International Edition)
Volume
61
Issue
38
DOI
doi:10.1002/anie.202207640
Link
https://onlinelibrary.wiley.com/doi/10.1002/anie.202207640

Funding

Sponsorship
NWO
Grant number
ERC-CoG 865175