Li, Jiaqing: Structure and function study of HCN1 channel by forming a lattice on the membrane. - Bonn, 2020. - Dissertation, Rheinische Friedrich-Wilhelms-Universität Bonn.
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author = {{Jiaqing Li}},
title = {Structure and function study of HCN1 channel by forming a lattice on the membrane},
school = {Rheinische Friedrich-Wilhelms-Universität Bonn},
year = 2020,
month = sep,

note = {Hyperpolarized-activated cyclic nucleotide-gated (HCN) channels play complex and diverse roles in the regulation of neuronal and network excitability. Aberrant expression, trafficking, and post-translational modifications of HCN1 channels contribute to the experimental and human epilepsy, yet the structural basis underpinning the altered function is still unknown. To unravel the structure-function relationship of HCN1 channels in the native cellular environment, we utilized an optimized intein system to cross-link channels into a structured lattice on the membrane. The successful forming of HCN1 lattice on the membrane was evidenced by split GFP reconstitution and functionally characterized by electrophysiological recording. We subsequently generated membrane sheets suitable for structural EM analysis directly from cells expressing protein lattices by unroofing techniques. The membrane sheets were then fluorescent- and immunolabeled with GFP nanobody conjugated quantum dot, which allowed the detection of protein lattice for correlative light and electron microscopy experiments. Compared to structural studies in membrane mimetic environments, this study provides invaluable opportunities for in situ structural determination of membrane protein in the native membrane environment.},
url = {}

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