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Novel insights into heme binding to hemoglobin

  • Marie-Therese Hopp ORCID logo , Dhruv Chetanbhai Rathod , Kristina Helena Winn , Shubhi Ambast and Diana Imhof ORCID logo EMAIL logo
Published/Copyright: August 31, 2022
Biological Chemistry
From the journal Biological Chemistry Volume 403 Issue 11-12

Abstract

Under hemolytic conditions, hemoglobin and subsequently heme are rapidly released, leading to the toxic effects characterizing diseases such as β-thalassemia and sickle cell disease. Herein, we provide evidence that human hemoglobin can bind heme in a transient fashion via surface-exposed sequence motifs. Following the synthesis of potential heme-binding motifs (HBMs) as peptides, their heme-binding capacity was investigated by UV–vis spectroscopy and ranked according to their binding affinity. Heme binding to human hemoglobin was subsequently studied by UV–vis and surface plasmon resonance (SPR) spectroscopy, revealing a heme-binding affinity in the sub- to micromolar range and a stoichiometry that clearly exceeds a 1:1 ratio. In silico molecular docking and simulation studies confirmed heme binding to the respective motifs in the β-chain of hemoglobin. Finally, the peroxidase-like activity of hemoglobin and the hemoglobin-heme complex was monitored, which indicated a much higher activity (>1800%) than other heme-peptide/protein complexes reported so far. The present study provides novel insights into the nature of intact hemoglobin concerning its transient interaction with heme, which suggests for the first time potential heme-scavenging properties of the protein at concomitant disassembly and, consequently, a potentiation of hemolysis and related processes.

Keywords: heme; heme-binding motifs; hemoglobin; hemolysis; methemoglobin
Corresponding author: Diana Imhof, Pharmaceutical Biochemistry and Bioanalytics, Pharmaceutical Institute, University of Bonn, An der Immenburg 4, D-53121 Bonn, Germany, E-mail:

Acknowledgments

The authors like to thank the University of Bonn for financial support (to D.I. and M.T.H.) and Matthias Geyer (University of Bonn) for providing access to the SPR device. In addition, technical assistance by Sabrina Linden and Cem Gündüz (both: University of Bonn) as well as supply of TFA by Solvay GmbH is gratefully acknowledged.

  1. Author contributions: D.I. designed the project. D.I. and M.T.H. planned the project. M.T.H. and K.H.W. performed the experiments. D.C.R. and S.A. carried out the computational studies. M.T.H., D.C.R., K.H.W., and S.A. analyzed the data. The manuscript was written by M.T.H. and D.I. and finalized through the contribution of all authors.

  2. Research funding: Financial support was received from the University of Bonn.

  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-2022-0188).

Received: 2022-05-24
Accepted: 2022-07-27
Published Online: 2022-08-31
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-2022-0188
Keywords for this article
heme; heme-binding motifs; hemoglobin; hemolysis; methemoglobin

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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