[en] [en] BACKGROUND AND PURPOSE: Venomous animals express numerous Kunitz-type peptides. The mambaquaretin-1 (MQ1) peptide identified from the Dendroaspis angusticeps venom is the most selective antagonist of the arginine-vasopressin V2 receptor (V2R) and the only unique Kunitz-type peptide active on a GPCR. We aimed to exploit other mamba venoms to enlarge the V2R-Kunitz peptide family and gain insight into the MQ1 molecular mode of action.
EXPERIMENTAL APPROACH: We used a bio-guided screening assay to identify novel MQs and placed them phylogenetically. MQs were produced by solid-phase peptide synthesis and characterized in vitro by binding and functional tests and in vivo by diuresis measurement in rats.
KEY RESULTS: Eight additional MQs were identified with nanomolar affinities for the V2R, all antagonists. MQs form a new subgroup in the Kunitz family, close to the V2R non-active dendrotoxins and to two V2R-active cobra toxins. Sequence comparison between active and non-active V2R Kunitz peptides highlighted five positions, among which four are involved in V2R interaction and belong to the two large MQ1 loops. We finally determined that eight positions, part of these two loops, interact with the V2R. The variant MQ1-K39A showed a higher affinity for the hV2R, but not for the rat V2R.
CONCLUSIONS AND IMPLICATIONS: A new function and mode of action is associated with the Kunitz peptides. The number of MQ1 residues involved in V2R binding is large and may explain its absolute selectivity. MQ1-K39A represents the first step in the improvement of the MQ1 design from a medicinal perspective.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Droctové, Laura; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Ciolek, Justyna; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Mendre, Christiane; Institut de Génomique Fonctionnelle, Université de Montpellier, CNRS, INSERM, Montpellier, France
Chorfa, Amélia; Institut de Génomique Fonctionnelle, Université de Montpellier, CNRS, INSERM, Montpellier, France
Huerta, Paola; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Carvalho, Chrystelle; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Gouin, Charlotte; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Lancien, Manon; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Stanajic-Petrovic, Goran; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Braco, Lorine; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Blanchet, Guillaume; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Upert, Gregory; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
De Pauw, Gregory; Laboratory of Mass Spectrometry, MolSys Research Unit, University of Liège, Liège, Belgium
Barbe, Peggy; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Keck, Mathilde; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Mourier, Gilles; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Mouillac, Bernard; Institut de Génomique Fonctionnelle, Université de Montpellier, CNRS, INSERM, Montpellier, France
Denis, Servent ; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
Rodríguez de la Vega, Ricardo C; Ecologie, Systematique Evolution, CNRS, AgroParisTech, Université Paris-Saclay, Orsay, France
Quinton, Loïc ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie biologique
Gilles, Nicolas; Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, Université Paris Saclay, CEA, INRAE, Gif-sur-Yvette, France
CEA - Commissariat à l'Énergie Atomique et aux Énergies Alternatives [FR] LNCC - Ligue Nationale Contre le Cancer [FR]
Funding text :
We thank Mathilde Gilles for her contribution in improving the quality of English language. This work was supported by the French Atomic and Alternative Energies and La Ligue contre le Cancer for financial support and for the Amelia Chorfa PhD programme funding.This work was supported by the French Atomic and Alternative Energies and for financial support and for the Amelia Chorfa PhD programme funding. La Ligue contre le Cancer
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