Molecular architecture of potassium chloride co-transporter KCC2

Autor: Eileen McCall, Anass Jawhari, Anders Gunnarsson, Niek Dekker, Ross A. Cardarelli, Stephen J. Moss, Kelly Garnier, Qi Wang, Stéphane Audebert, Tarek Z. Deeb, Stefan Geschwindner, Matthew P. Burnham, Patrick Schultz, David Baker, Alexandre Frechard, Igor Medina, Morgane Agez, Leslie C. Conway, Nicholas J. Brandon
Přispěvatelé: Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Epilepsie et ischémie cérébrale, Université de la Méditerranée - Aix-Marseille 2-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Library of JiLin Agriculture University, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU)
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
[SDV]Life Sciences [q-bio]
lcsh:Medicine
[SDV.CAN]Life Sciences [q-bio]/Cancer
Chloride
Article
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Genomics [q-bio.GN]
Calixarene
medicine
Surface plasmon resonance
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biochemistry [q-bio.BM]
lcsh:Science
ComputingMilieux_MISCELLANEOUS
Multidisciplinary
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Structural Biology [q-bio.BM]
lcsh:R
Transporter
Potassium channel
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biomolecules [q-bio.BM]
030104 developmental biology
Monomer
chemistry
Biophysics
lcsh:Q
Linker
030217 neurology & neurosurgery
Function (biology)
medicine.drug
Zdroj: Scientific Reports, Vol 7, Iss 1, Pp 1-14 (2017)
Scientific Reports
Scientific Reports, Nature Publishing Group, 2017, 7 (1), ⟨10.1038/s41598-017-15739-1⟩
Scientific Reports, 2017, 7 (1), ⟨10.1038/s41598-017-15739-1⟩
ISSN: 2045-2322
DOI: 10.1038/s41598-017-15739-1
Popis: KCC2 is a neuron specific K+-Cl− co-transporter that controls neuronal chloride homeostasis, and is critically involved in many neurological diseases including brain trauma, epilepsies, autism and schizophrenia. Despite significant accumulating data on the biology and electrophysiological properties of KCC2, structure-function relationships remain poorly understood. Here we used calixarene detergent to solubilize and purify wild-type non-aggregated and homogenous KCC2. Specific binding of inhibitor compound VU0463271 was demonstrated using surface plasmon resonance (SPR). Mass spectrometry revealed glycosylations and phosphorylations as expected from functional KCC2. We show by electron microscopy (EM) that KCC2 exists as monomers and dimers in solution. Monomers are organized into “head” and “core” domains connected by a flexible “linker”. Dimers are asymmetrical and display a bent “S-shape” architecture made of four distinct domains and a flexible dimerization interface. Chemical crosslinking in reducing conditions shows that disulfide bridges are involved in KCC2 dimerization. Moreover, we show that adding a tag to the C-terminus is detrimental to KCC2 function. We postulate that the conserved KCC2 C-ter may be at the interface of dimerization. Taken together, our findings highlight the flexible multi-domain structure of KCC2 with variable anchoring points at the dimerization interface and an important C-ter extremity providing the first in-depth functional architecture of KCC2.
Databáze: OpenAIRE
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