Directional K+ channel insertion in a single phospholipid bilayer: Neutron reflectometry and electrophysiology in the joint exploration of a model membrane functional platform

Biochimica Et Biophysica Acta. General Subjects
V RondelliL Cantu'

Abstract

We investigated the insertion of small potassium (K+) channel proteins (KcvMA-1D and KcvNTS) into model membranes and the lipid-protein structural interference, combining neutron reflectometry and electrophysiology. Neutron reflectometry experiments showed how the transverse structure and mechanical properties of the bilayer were modified, upon insertion of the proteins in single model-membranes, either supported on solid substrate or floating. Parallel electrophysiology experiments were performed on the same channels reconstituted in free-standing planar lipid bilayers, of both typical composition and matched to the neutron reflectometry experiment, assessing their electrical features. Functional and structural results converge in detecting that the proteins, conical in shape, insert with a directionality, cytosolic side first. Our work addresses the powerful combination of the two experimental approaches. We show here that membrane structure spectroscopy and ion channel electrophysiology can become synergistic tools in the analysis of structural-functional properties of biomimetic complex environment.

Citations

Aug 1, 2019·International Journal of Molecular Sciences·Valeria RondelliPaola Brocca
Jul 21, 2020·Journal of Applied Crystallography·Frank HeinrichMathias Lösche
Nov 25, 2020·Cells·Anja J EngelGerhard Thiel
Jan 14, 2021·The Journal of General Physiology·Laura-Marie WintersteinOliver Rauh
May 15, 2021·Frontiers in Chemistry·Jacob J KinnunJohn Katsaras

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