title: Characterization of the direct interaction between KcsA-Kv1.3 and its inhibitors.
abstract: Integral membrane proteins (IMPs) are of therapeutic interest and are targeted by a majority of approved drugs. It's difficult to express, purify, and maintain the functional conformation of IMPs. Nanodisc presents a reliable method to solubilize and stabilize IMPs in detergent-free condition. In this study, we demonstrate the assembly and purification of a chimeric ion channel, KcsA-Kv1.3 Nanodisc. We further detail biophysical analysis of the assembled Nanodisc using analytical ultracentrifugation (AUC), surface plasmon resonance (SPR), and back scattering interferometry (BSI). AUC is employed to determine the molecular composition of the empty and KcsA-Kv1.3 Nanodisc. Combination of SPR and BSI overcomes each other's limitation and provides insight of equilibrium binding properties of peptide and small molecule ligands to KcsA-Kv1.3.
authors.affiliation: Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA 91320, United States.
authors.forename: Han
authors.initials: H
authors.lastname: Xu
authors.affiliation: Therapeutic Discovery, Amgen Inc., Seattle, WA 98119, United States.
authors.forename: John J
authors.initials: JJ
authors.lastname: Hill
authors.affiliation: Therapeutic Discovery, Amgen Inc., Cambridge, MA 02142, United States.
authors.forename: Klaus
authors.initials: K
authors.lastname: Michelsen
authors.affiliation: Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA 91320, United States.
authors.forename: Harvey
authors.initials: H
authors.lastname: Yamane
authors.affiliation: Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA 91320, United States.
authors.forename: Robert J M
authors.initials: RJ
authors.lastname: Kurzeja
authors.affiliation: Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA 91320, United States.
authors.forename: Tony
authors.initials: T
authors.lastname: Tam
authors.affiliation: Molecular Sensing Inc., Nashville, TN 37203, United States.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Isaacs
authors.affiliation: Molecular Sensing Inc., Nashville, TN 37203, United States.
authors.forename: Fei
authors.initials: F
authors.lastname: Shen
authors.affiliation: Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA 91320, United States.
authors.forename: Philip
authors.initials: P
authors.lastname: Tagari
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D001665:Binding Sites; D051664:Kv1.3 Potassium Channel / Q000037:antagonists & inhibitors / Q000737:chemistry*; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000138:chemical synthesis / Q000648:ultrastructure; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure*; D026902:Potassium Channel Blockers / Q000737:chemistry*; D015221:Potassium Channels / Q000737:chemistry*; D011485:Protein Binding
chemical_list: D001426:Bacterial Proteins; D051664:Kv1.3 Potassium Channel; D046912:Multiprotein Complexes; D026902:Potassium Channel Blockers; D015221:Potassium Channels; C107790:prokaryotic potassium channel
keywords: Ion channel ligand binding; KcsA-Kv1.3; Nanodisc