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Related Experiment Video

Updated: Dec 9, 2025

Selection of Plasmodium falciparum Parasites for Cytoadhesion to Human Brain Endothelial Cells

Selection of Plasmodium falciparum Parasites for Cytoadhesion to Human Brain Endothelial Cells

Published on: January 3, 2012

Designed Parasite-Selective Rhomboid Inhibitors Block Invasion and Clear Blood-Stage Malaria.

Shiv Gandhi¹, Rosanna P Baker¹, Sangwoo Cho¹

¹Department of Molecular Biology & Genetics, Johns Hopkins University School of Medicine, Room 507 PCTB, 725 North Wolfe Street, Baltimore, MD 21205, USA.

Cell Chemical Biology

|September 5, 2020

Summary

Researchers developed parasite-selective rhomboid inhibitors targeting malaria. This strategy blocks host-cell invasion and clears parasitemia, offering a new approach for treating malaria by inhibiting essential rhomboid proteolysis.

Keywords:

Plasmodium Ras-converting enzyme Toxoplasma apicomplexan parasites malaria presenilin regulated intramembrane proteolysis rhomboid protease serine protease site-2 protease

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Area of Science:

Biochemistry
Parasitology
Drug Discovery

Background:

Rhomboid intramembrane proteases are crucial in various pathophysiological processes.
Targeting rhomboid proteases for disease intervention has remained elusive.
The malaria parasite's rhomboid PfROM4 exhibits atypical substrate specificity.

Purpose of the Study:

To decode the substrate specificity of PfROM4.
To engineer parasite-selective rhomboid inhibitors.
To establish a strategy for targeting rhomboid proteolysis in malaria.

Main Methods:

Investigated substrate specificity through steric exclusion.
Engineered an optimal substrate sequence to enhance proteolysis.
Utilized high-resolution structural analysis to discover boronate inhibitors.
Screened compound libraries for orthogonal inhibitors.
Performed ultrastructural analysis on malaria-infected cultures.

Main Results:

PfROM4's specificity arises from steric exclusion, preventing cross-reactivity with human rhomboids.
Engineered substrates enhanced proteolysis over 10-fold.
Boronate inhibitors demonstrated over 100-fold enhancement.
Developed parasite-selective peptide boronate and alpha-ketoamide inhibitors.
Single-dose treatment blocked host-cell invasion and cleared parasitemia in vitro.

Conclusions:

A strategy for designing parasite-selective rhomboid inhibitors was established.
PfROM4 is a druggable target in non-motile malaria parasites.
Inhibition of rhomboid proteolysis offers a novel therapeutic approach for malaria.