Individual cells traffic the Vasopressin 2 Receptor to their cell surface with different success Eline J. Koers 1,2,* , Bradley A. Morgan 1,3,4 , Iain B. Styles 1,3 and Dmitry B. Veprintsev 1,2,* 1 Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK 2 Division of Physiology, Pharmacology & Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, NG7 2UH, UK 3 School of Computer Science, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK and The Alan Turing Institute, London, NW1 2DB, UK 4 MRC IMPACT Doctoral Training Programme, Universities of Birmingham, Leicester and Nottingham, Midlands, UK *correspondence should be addressed to Eline.Koers@nottingham.ac.uk and Dmitry.Veprintsev@nottingham.ac.uk. Summary G protein coupled receptors (GPCRs) translate the actions of hormones and neurotransmitters into intracellular signalling events. Mutations in GPCRs can prevent their correct expression and trafficking to the cell surface and cause disease. Single cell subcellular localisation measurements reveal that while some cells appear to traffic the majority of the vasopressin 2 receptor (V2R) molecules to the cell surface, others retain a greater number of receptors in the ER or have approximately equal distribution. Mutations in the V2R affect the proportion of cells able to send this GPCR to their cell surface but surprisingly they do not prevent all cells from correctly trafficking the mutant receptors. These findings reveal the potential for rescue of mutant receptor cell surface expression by pharmacological manipulation of the GPCR folding and trafficking machinery. Introduction G protein coupled receptors (GPCRs) coordinate functions of multicellular organisms by sensing hormones, neurotransmitters or cytokines that circulate around the body and inducing intracellular responses. Missense mutations in these receptors often cause loss-of-function and as such mutations are linked to 55 monogenic diseases (Schöneberg, Liebscher et al. 2020). Missense mutations in GPCRs can lead to a change or loss of ligand binding and intracellular signalling (i.e. function), but the majority of mutations causing disease are located outside of the key functional sites -i.e. ligand binding pocket, G protein and β- arrestin binding sites- (Hauser, Chavali et al. 2017) and hence are more likely to affect the folding, structure or stability of the GPCR. . CC-BY 4.0 International license made available under a (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is The copyright holder for this preprint this version posted August 9, 2021. ; https://doi.org/10.1101/2021.08.09.455709 doi: bioRxiv preprint