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Purification and membrane interactions of human KCNQ1100-370 potassium ion channel.
Dixit, Gunjan; Stowe, Rebecca B; Bates, Alison; Jaycox, Colleen K; Escobar, Jorge R; Harding, Benjamin D; Drew, Daniel L; New, Christopher P; Sahu, Indra D; Edelmann, Richard E; Dabney-Smith, Carole; Sanders, Charles R; Lorigan, Gary A.
Affiliation
  • Dixit G; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA; Cell, Molecular and Structural Biology Program, Department of Chemistry & Biochemistry, Miami University, Oxford, OH 45056, USA.
  • Stowe RB; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA.
  • Bates A; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA.
  • Jaycox CK; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA.
  • Escobar JR; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA; Cell, Molecular and Structural Biology Program, Department of Chemistry & Biochemistry, Miami University, Oxford, OH 45056, USA.
  • Harding BD; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA.
  • Drew DL; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA.
  • New CP; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA; Cell, Molecular and Structural Biology Program, Department of Chemistry & Biochemistry, Miami University, Oxford, OH 45056, USA.
  • Sahu ID; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA.
  • Edelmann RE; Center for Advanced Microscopy and Imaging, Miami University, Oxford, OH 45056, USA.
  • Dabney-Smith C; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA; Cell, Molecular and Structural Biology Program, Department of Chemistry & Biochemistry, Miami University, Oxford, OH 45056, USA.
  • Sanders CR; Department of Biochemistry and Center for Structural Biology, Vanderbilt University, Nashville, TN 37240, USA.
  • Lorigan GA; Department of Chemistry and Biochemistry, Miami University, 651 E. High Street, Oxford, OH 45056, USA; Cell, Molecular and Structural Biology Program, Department of Chemistry & Biochemistry, Miami University, Oxford, OH 45056, USA. Electronic address: gary.lorigan@miamioh.edu.
Biochim Biophys Acta Biomembr ; 1864(11): 184010, 2022 11 01.
Article in En | MEDLINE | ID: mdl-35870481
KCNQ1 (Kv7.1 or KvLQT1) is a voltage-gated potassium ion channel that is involved in the ventricular repolarization following an action potential in the heart. It forms a complex with KCNE1 in the heart and is the pore forming subunit of slow delayed rectifier potassium current (Iks). Mutations in KCNQ1, leading to a dysfunctional channel or loss of activity have been implicated in a cardiac disorder, long QT syndrome. In this study, we report the overexpression, purification, biochemical characterization of human KCNQ1100-370, and lipid bilayer dynamics upon interaction with KCNQ1100-370. The recombinant human KCNQ1 was expressed in Escherichia coli and purified into n-dodecylphosphocholine (DPC) micelles. The purified KCNQ1100-370 was biochemically characterized by SDS-PAGE electrophoresis, western blot and nano-LC-MS/MS to confirm the identity of the protein. Circular dichroism (CD) spectroscopy was utilized to confirm the secondary structure of purified protein in vesicles. Furthermore, 31P and 2H solid-state NMR spectroscopy in DPPC/POPC/POPG vesicles (MLVs) indicated a direct interaction between KCNQ100-370 and the phospholipid head groups. Finally, a visual inspection of KCNQ1100-370 incorporated into MLVs was confirmed by transmission electron microscopy (TEM). The findings of this study provide avenues for future structural studies of the human KCNQ1 ion channel to have an in depth understanding of its structure-function relationship.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Long QT Syndrome / Potassium Channels, Voltage-Gated Limits: Humans Language: En Journal: Biochim Biophys Acta Biomembr Year: 2022 Document type: Article Affiliation country: Estados Unidos Country of publication: Países Bajos
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Long QT Syndrome / Potassium Channels, Voltage-Gated Limits: Humans Language: En Journal: Biochim Biophys Acta Biomembr Year: 2022 Document type: Article Affiliation country: Estados Unidos Country of publication: Países Bajos