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Related Experiment Video

Updated: Jul 5, 2025

Method for Identifying Small Molecule Inhibitors of the Protein-protein Interaction Between HCN1 and TRIP8b
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Hemin competitively inhibits HSPA8 ATPase activity mitigating its foldase function.

Alok Kumar Pandey1, Vishal Trivedi1

  • 1Malaria Research Group, Department of Bioscience and Bioengineering, Indian Institute of Technology-Guwahati, Guwahati, 781039, Assam, India.

Archives of Biochemistry and Biophysics
|January 12, 2024
PubMed
Summary

High hemin levels inhibit the chaperone HSPA8

Keywords:
Competitive-inhibitionFoldaseHSPA8HeminMalariaOxidative stress

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Biology

Background:

  • Hemolysis leads to elevated hemin levels, contributing to severe malaria pathology.
  • Hemin's structure facilitates protein binding and functional modification.
  • HSPA8 is a chaperone protein involved in oxidative stress protection via protein refolding.

Purpose of the Study:

  • To investigate the interaction between hemin and the chaperone HSPA8.
  • To determine how hemin affects HSPA8's chaperone activity and ATP hydrolysis.
  • To elucidate the mechanism of HSPA8 inhibition by hemin.

Main Methods:

  • ATPase activity assays
  • Circular Dichroism (CD) spectroscopy
  • Gel filtration
  • In-silico molecular docking
  • Isothermal titration calorimetry (ITC)

Main Results:

  • Hemin competitively inhibits HSPA8's ATP hydrolysis and foldase function.
  • Hemin does not alter HSPA8's structural integrity.
  • Hemin binding reduces ATP affinity for HSPA8 by 22-fold, hindering chaperone activity.
  • ATP cannot displace hemin from the HSPA8 ATP-binding pocket.

Conclusions:

  • Hemin inhibits HSPA8's cytoprotective function by competing with ATP binding.
  • This inhibition may contribute to cellular damage in hemolytic diseases like severe malaria.
  • Hemin's interference with chaperone function represents a novel pathogenic mechanism.