RT Journal Article SR Electronic T1 A synthetic method to assay polycystin channel biophysics JF bioRxiv FD Cold Spring Harbor Laboratory SP 2024.05.06.592666 DO 10.1101/2024.05.06.592666 A1 Larmore, Megan A1 Esarte Palomero, Orhi A1 Kamat, Neha P. A1 DeCaen, Paul G. YR 2024 UL http://biorxiv.org/content/early/2024/08/10/2024.05.06.592666.abstract AB Ion channels are biological transistors that control ionic flux across cell membranes to regulate electrical transmission and signal transduction. They are found in all biological membranes and their conductive state kinetics are frequently disrupted in human diseases. Organelle ion channels are among the most resistant to functional and pharmacological interrogation. Traditional channel protein reconstitution methods rely upon exogenous expression and/or purification from endogenous cellular sources which are frequently contaminated by resident ionophores. Here we describe a fully synthetic method to assay functional properties of polycystin channels that natively traffic to primary cilia and endoplasmic reticulum organelles. Using this method, we characterize their oligomeric assembly, membrane integration, orientation and conductance while comparing these results to their endogenous channel properties. Outcomes define a novel synthetic approach that can be applied broadly to investigate channels resistant to biophysical analysis and pharmacological characterization.Competing Interest StatementThe authors have declared no competing interest.