TY - JOUR
T1 - Live-Cell Imaging of Sterculic Acid—a Naturally Occurring 1,2-Cyclopropene Fatty Acid—by Bioorthogonal Reaction with Turn-On Tetrazine-Fluorophore Conjugates**
AU - Bertheussen, Kristine
AU - van de Plassche, Merel
AU - Bakkum, Thomas
AU - Gagestein, Berend
AU - Ttofi, Iakovia
AU - Sarris, Alexi J.C.
AU - Overkleeft, Herman S.
AU - van der Stelt, Mario
AU - van Kasteren, Sander I.
N1 - Funding Information: We would like to acknowledge Ken Rock for his kind gift of the DC2.4 cell line. We would also kindly like to thank the Institute for Chemical Immunology (NWO-Zwaartekracht Grant) and the European Research Council (ERC-CoG 865175) for funding the work described here. Funding Information: We would like to acknowledge Ken Rock for his kind gift of the DC2.4 cell line. We would also kindly like to thank the Institute for Chemical Immunology (NWO‐Zwaartekracht Grant) and the European Research Council (ERC‐CoG 865175) for funding the work described here. Publisher Copyright: © 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.
PY - 2022/9/19
Y1 - 2022/9/19
N2 - 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 by multimodal confocal imaging.
AB - 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 by multimodal confocal imaging.
KW - Click Chemistry
KW - Cyclopropene
KW - Inverse Electron-Demand Diels–Alder Reaction
KW - Lipids
KW - Sterculic Acid
UR - http://www.scopus.com/inward/record.url?scp=85135700159&partnerID=8YFLogxK
U2 - https://doi.org/10.1002/anie.202207640
DO - https://doi.org/10.1002/anie.202207640
M3 - Article
C2 - 35838324
SN - 1433-7851
VL - 61
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 38
M1 - e202207640
ER -