Nonperturbative fluorogenic labeling of immunophilins enables the wash-free detection of immunosuppressants

Bertolini, Marco, Mendive-Tapia, Lorena, Ghashghaei, Ouldouz, Reese, Abigail, Lochenie, Charles, Schoepf, Anna M., Sintes, Miquel, Tokarczyk, Karolina, Nare, Zandile, Scott, Andrew D., Knight, Stephen R., Aithal, Advait R., Sachdeva, Amit, Lavilla, Rodolfo and Vendrell, Marc (2024) Nonperturbative fluorogenic labeling of immunophilins enables the wash-free detection of immunosuppressants. ACS Central Science, 10 (5). 969–977. ISSN 2374-7943

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Abstract

Immunosuppressants are clinically approved drugs to treat the potential rejection of transplanted organs and require frequent monitoring due to their narrow therapeutic window. Immunophilins are small proteins that bind immunosuppressants with high affinity, yet there are no examples of fluorogenic immunophilins and their potential application as optical biosensors for immunosuppressive drugs in clinical biosamples. In the present work, we designed novel diazonium BODIPY salts for the site-specific labeling of tyrosine residues in peptides via solid-phase synthesis as well as for late-stage functionalization of whole recombinant proteins. After the optimization of a straightforward one-step labeling procedure for immunophilins PPIA and FKBP12, we demonstrated the application of a fluorogenic analogue of FKBP12 for the selective detection of the immunosuppressant drug tacrolimus, including experiments in urine samples from patients with functioning renal transplants. This chemical methodology opens new avenues to rationally design wash-free immunophilin-based biosensors for rapid therapeutic drug monitoring.

Item Type:	Article
Additional Information:	Acknowledgments: L.M.-T. acknowledges funding from the Wellcome Trust Institutional Strategic Support Fund (ISSF) at the University of Edinburgh. O.G. acknowledges support through the Eurolife Postdoc Mobility and Knowledge Exchange Program. A.M.S. thanks the Austrian Science Fund (FWF) for the project J4541-N. A.R.A. is funded by the NRP-BBSRC-Doctoral Training Programme. S.R.K. acknowledges funding from a Kidney Research UK Stoneygate Startup award (KS_SU_002_20221129). A.S. acknowledges funding from the University of East Anglia. R.L. acknowledges the Ministerio de Ciencia e Innovación (Spain), the European Regional Development Fund (EDFR) (PID2022-139180OB-I00), and Generalitat de Catalunya (Research Quality groups SGR-DGR 00357). M.V. acknowledges funds from an ERC Consolidator Grant (DYNAFLUORS, 771443). The authors acknowledge the EdinOmics research facility (RRID: SCR_021838, University of Edinburgh) for carrying out the mass spectrometry analyses of the proteins. This project has received funding from the European Union’s Horizon 2020 Research and Innovation Program under the Marie Sklodowska-Curie grant agreement (956477). For open access, the authors applied a CC-BY public copyright license to any Author Accepted Manuscript version arising from this submission. The authors acknowledge BioRender.com for assistance with figure creation and Silvia Dotto for the graphical design and creation of the cover art.
Faculty \ School:	Faculty of Science > School of Chemistry, Pharmacy and Pharmacology Faculty of Science
UEA Research Groups:	Faculty of Science > Research Groups > Chemistry of Life Processes
Depositing User:	LivePure Connector
Date Deposited:	04 Oct 2024 08:30
Last Modified:	16 Oct 2024 00:05
URI:	https://ueaeprints.uea.ac.uk/id/eprint/96878
DOI:	10.1021/acscentsci.3c01590

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