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Extracellular hemin is a reverse use-dependent gating modifier of cardiac voltage-gated Na+ channels

  • Guido Gessner , Mahdi Jamili , Pascal Tomczyk , Dirk Menche , Roland Schönherr , Toshinori Hoshi and Stefan H. Heinemann ORCID logo EMAIL logo
Published/Copyright: August 29, 2022
Biological Chemistry
From the journal Biological Chemistry Volume 403 Issue 11-12

Abstract

Heme (Fe2+-protoporphyrin IX) is a well-known protein prosthetic group; however, heme and hemin (Fe3+-protoporphyrin IX) are also increasingly viewed as signaling molecules. Among the signaling targets are numerous ion channels, with intracellular-facing heme-binding sites modulated by heme and hemin in the sub-µM range. Much less is known about extracellular hemin, which is expected to be more abundant, in particular after hemolytic insults. Here we show that the human cardiac voltage-gated sodium channel hNaV1.5 is potently inhibited by extracellular hemin (IC50 ≈ 80 nM), while heme, dimethylhemin, and protoporphyrin IX are ineffective. Hemin is selective for hNaV1.5 channels: hNaV1.2, hNaV1.4, hNaV1.7, and hNaV1.8 are insensitive to 1 µM hemin. Using domain chimeras of hNaV1.5 and rat rNaV1.2, domain II was identified as the critical determinant. Mutation N803G in the domain II S3/S4 linker largely diminished the impact of hemin on the cardiac channel. This profile is reminiscent of the interaction of some peptide voltage-sensor toxins with NaV channels. In line with a mechanism of select gating modifiers, the impact of hemin on NaV1.5 channels is reversely use dependent, compatible with an interaction of hemin and the voltage sensor of domain II. Extracellular hemin thus has potential to modulate the cardiac function.

Keywords: heme; NaV1.5; SCN5A ; sodium channel; voltage sensor
Corresponding author: Stefan H. Heinemann, Department of Biophysics, Center for Molecular Biomedicine, Friedrich Schiller University Jena and Jena University Hospital, Hans-Knöll-Straße 2, D-07745 Jena, Germany, E-mail:

Funding source: National Institutes of Health

Award Identifier / Grant number: GM121375

Funding source: Deutsche Forschungsgemeinschaft

Award Identifier / Grant number: HE2993/18-1

Acknowledgments

We thank Prof. Enrico Leipold (University of Lübeck, Germany) for constructing hNaV1.8c4.

  1. Author contributions: G.G.: study design, molecular biology of expression constructs, electrophysiological recordings, data analysis; M.J.: electrophysiological recordings, data analysis; P.T., D.M.: synthesis of dimethylhemin; R.S.: molecular biology of expression constructs; T.H.: structural modeling, data review, writing; S.H.H.: study design, data analysis, writing. All authors contributed to the editing of the manuscript.

  2. Research funding: Support by the German Research Foundation (DFG, SHH: HE2993/18-1) and the National Institutes of Health (NIH, TH: GM121375).

  3. Conflict of interest statement: The authors declare no conflicts of interest regarding this article.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/hsz-2022-0194).

Received: 2022-06-01
Accepted: 2022-08-10
Published Online: 2022-08-29
Published in Print: 2022-11-25

© 2022 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Heme research – the past, the present and the future
  3. A primer on heme biosynthesis
  4. New roles for GAPDH, Hsp90, and NO in regulating heme allocation and hemeprotein function in mammals
  5. The role of host heme in bacterial infection
  6. Signal transduction mechanisms in heme-based globin-coupled oxygen sensors with a focus on a histidine kinase (AfGcHK) and a diguanylate cyclase (YddV or EcDosC)
  7. Heme delivery to heme oxygenase-2 involves glyceraldehyde-3-phosphate dehydrogenase
  8. Novel insights into heme binding to hemoglobin
  9. Extracellular hemin is a reverse use-dependent gating modifier of cardiac voltage-gated Na+ channels
  10. Assessment of the breadth of binding promiscuity of heme towards human proteins
  11. Determination of free heme in stored red blood cells with an apo-horseradish peroxidase-based assay
  12. Comparative studies of soluble and immobilized Fe(III) heme-peptide complexes as alternative heterogeneous biocatalysts
https://doi.org/10.1515/hsz-2022-0194
Keywords for this article
heme; NaV1.5; SCN5A; sodium channel; voltage sensor

Articles in the same Issue

  1. Frontmatter
  2. Heme research – the past, the present and the future
  3. A primer on heme biosynthesis
  4. New roles for GAPDH, Hsp90, and NO in regulating heme allocation and hemeprotein function in mammals
  5. The role of host heme in bacterial infection
  6. Signal transduction mechanisms in heme-based globin-coupled oxygen sensors with a focus on a histidine kinase (AfGcHK) and a diguanylate cyclase (YddV or EcDosC)
  7. Heme delivery to heme oxygenase-2 involves glyceraldehyde-3-phosphate dehydrogenase
  8. Novel insights into heme binding to hemoglobin
  9. Extracellular hemin is a reverse use-dependent gating modifier of cardiac voltage-gated Na+ channels
  10. Assessment of the breadth of binding promiscuity of heme towards human proteins
  11. Determination of free heme in stored red blood cells with an apo-horseradish peroxidase-based assay
  12. Comparative studies of soluble and immobilized Fe(III) heme-peptide complexes as alternative heterogeneous biocatalysts
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