2. Academic Validation
  3. Role of human Hv1 channels in sperm capacitation and white blood cell respiratory burst established by a designed peptide inhibitor

Role of human Hv1 channels in sperm capacitation and white blood cell respiratory burst established by a designed peptide inhibitor

  • Proc Natl Acad Sci U S A. 2018 Dec 11;115(50):E11847-E11856. doi: 10.1073/pnas.1816189115.
Ruiming Zhao 1 2 3,  Kelleigh Kennedy 1,  Gerardo A De Blas 4,  Gerardo Orta 5,  Martín A Pavarotti 4,  Rodolfo J Arias 4,  José Luis de la Vega-Beltrán 5,  Qufei Li 6,  Hui Dai 1 2 3,  Eduardo Perozo 6,  Luis S Mayorga 4,  Alberto Darszon 5,  Steve A N Goldstein 7 2 3
Affiliations

Affiliations

  • 1 Department of Biochemistry, Brandeis University, Waltham, MA 02453.
  • 2 Department of Pediatrics, Loyola University Chicago Stritch School of Medicine, Maywood, IL 60153.
  • 3 Department of Cell and Molecular Physiology, Loyola University Chicago Stritch School of Medicine, Maywood, IL 60153.
  • 4 Instituto de Histología y Embriología de Mendoza (Consejo Nacional de Investigaciones Científicas y Técnicas-Universidad Nacional de Cuyo), School of Medicine, National University of Cuyo, Mendoza CP 5500, Argentina.
  • 5 Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, 62250 Morelos, México.
  • 6 Department of Biochemistry and Molecular Biology, Gordon Center for Integrative Science, University of Chicago, Chicago, IL 60637.
  • 7 Department of Biochemistry, Brandeis University, Waltham, MA 02453; sgoldstein3@luc.edu.
PMID: 30478045 DOI: 10.1073/pnas.1816189115
Abstract

Using a de novo peptide inhibitor, Corza6 (C6), we demonstrate that the human voltage-gated proton channel (hHv1) is the main pathway for H+ efflux that allows capacitation in sperm and permits sustained Reactive Oxygen Species (ROS) production in white blood cells (WBCs). C6 was identified by a phage-display strategy whereby ∼1 million novel Peptides were fabricated on an inhibitor cysteine knot (ICK) scaffold and sorting on purified hHv1 protein. Two C6 Peptides bind to each dimeric channel, one on the S3-S4 loop of each voltage sensor domain (VSD). Binding is cooperative with an equilibrium affinity (Kd) of ∼1 nM at -50 mV. As expected for a VSD-directed toxin, C6 inhibits by shifting hHv1 activation to more positive voltages, slowing opening and speeding closure, effects that diminish with membrane depolarization.

Keywords

C6; ICK; TIRF; proton channel; venom.

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