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Heme delivery to heme oxygenase-2 involves glyceraldehyde-3-phosphate dehydrogenase

  • Yue Dai ORCID logo , Angela S. Fleischhacker ORCID logo , Liu Liu ORCID logo , Sara Fayad , Amanda L. Gunawan ORCID logo , Dennis J. Stuehr ORCID logo and Stephen W. Ragsdale ORCID logo EMAIL logo
Published/Copyright: October 28, 2022
Biological Chemistry
From the journal Biological Chemistry Volume 403 Issue 11-12

Abstract

Heme regulatory motifs (HRMs) are found in a variety of proteins with diverse biological functions. In heme oxygenase-2 (HO2), heme binds to the HRMs and is readily transferred to the catalytic site in the core of the protein. To further define this heme transfer mechanism, we evaluated the ability of GAPDH, a known heme chaperone, to transfer heme to the HRMs and/or the catalytic core of HO2. Our results indicate GAPDH and HO2 form a complex in vitro. We have followed heme insertion at both sites by fluorescence quenching in HEK293 cells with HO2 reporter constructs. Upon mutation of residues essential for heme binding at each site in our reporter construct, we found that HO2 binds heme at the core and the HRMs in live cells and that heme delivery to HO2 is dependent on the presence of GAPDH that is competent for heme binding. In sum, GAPDH is involved in heme delivery to HO2 but, surprisingly, not to a specific site on HO2. Our results thus emphasize the importance of heme binding to both the core and the HRMs and the interplay of HO2 with the heme pool via GAPDH to maintain cellular heme homeostasis.

Keywords: chaperone; GAPDH; glyceraldehyde-3-phosphate dehydrogenase; heme oxygenase-2; heme regulatory motifs; heme trafficking
Corresponding author: Stephen W. Ragsdale, Department of Biological Chemistry, University of Michigan Medical School, 1150 W. Medical Center Dr., 5301 MSRB III, Ann Arbor, MI 48109-0606, E-mail:
Yue Dai and Angela S. Fleischhacker contributed equally to this work.

Funding source: National Institute of General Medical Sciences

Award Identifier / Grant number: R01GM130624

Award Identifier / Grant number: R35-GM141758

Funding source: National Institutes of Health

Award Identifier / Grant number: Unassigned

Acknowledgements

We thank Prof. Zhan Chen and lab members for assistance with CD experiments, Dr. Anindita Sarkar for helpful discussion and valuable feedback, and Claire Maiocco for assistance with method development.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: This work was supported by National Institutes of Health Grants R01GM130624 (to D.J.S.), R01-GM123513 (to S.W.R.) and R35-GM141758 (to S.W.R.).

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

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Received: 2022-07-12
Accepted: 2022-10-10
Published Online: 2022-10-28
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-2022-0230
Keywords for this article
chaperone; GAPDH; glyceraldehyde-3-phosphate dehydrogenase; heme oxygenase-2; heme regulatory motifs; heme trafficking

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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