Back to Search Start Over

Cross-Linking Furan-Modified Kisspeptin-10 to the KISS Receptor

Authors :
Annemieke Madder
Massimiliano Beltramo
Christophe Ampe
Willem Vannecke
Marleen Van Troys
Organic and Biomimetic Chemistry Research Group
Department of Biochemistry, Faculty of Medicine and Health Sciences
Universiteit Gent = Ghent University [Belgium] (UGENT)
Physiologie de la reproduction et des comportements [Nouzilly] (PRC)
Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)
BOF-UGent under grant agreement number BOF15/DOC_V/404
Research Foundation Flanders under grant agreement number G048516N
Region Centre Val de Loire council (grant Capriss)
French Agence National de la Recherche (grant ANR-15-CE20-0015- 01)
Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)
Ghent University [Belgium] (UGENT)
Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA)
Source :
ACS Chemical Biology, ACS Chemical Biology, American Chemical Society, 2017, 12 (8), pp.2191-2200. ⟨10.1021/acschembio.7b00396⟩
Publication Year :
2017

Abstract

Chemical cross-linking is well-established for investigating protein–protein interactions. Traditionally, photo cross-linking is used but is associated with problems of selectivity and UV toxicity in a biological context. We here describe, with live cells and under normal growth conditions, selective cross-linking of a furan-modified peptide ligand to its membrane-presented receptor with zero toxicity, high efficiency, and spatio-specificity. Furan-modified kisspeptin-10 is covalently coupled to its glycosylated membrane receptor, GPR54(KISS1R). This newly expands the applicability of furan-mediated cross-linking not only to protein–protein cross-linking but also to cross-linking in situ. Moreover, in our earlier reports on nucleic acid interstrand cross-linking, furan activation required external triggers of oxidation (via addition of N-bromo succinimide or singlet oxygen). In contrast, we here show, for multiple cell lines, the spontaneous endogenous oxidation of the furan moiety with concurrent selective cross-link formation. We propose that reactive oxygen species produced by NADPH oxidase (NOX) enzymes form the cellular source establishing furan oxidation.

Details

ISSN :
15548937 and 15548929
Volume :
12
Issue :
8
Database :
OpenAIRE
Journal :
ACS chemical biology
Accession number :
edsair.doi.dedup.....3f1411f13e54f0a62587f2b8a91c541c
Full Text :
https://doi.org/10.1021/acschembio.7b00396⟩