Structure of the Plasmodium falciparum M17 aminopeptidase and significance for the design of drugs targeting the neutral exopeptidases.

Abstract

Current therapeutics and prophylactics for malaria are under severe challenge as a result of the rapid emergence of drug-resistant parasites. The human malaria parasite Plasmodium falciparum expresses two neutral aminopeptidases, PfA-M1 and PfA-M17, which function in regulating the intracellular pool of amino acids required for growth and development inside the red blood cell. These enzymes are essential for parasite viability and are validated therapeutic targets. We previously reported the X-ray crystal structure of the monomeric PfA-M1 and proposed a mechanism for substrate entry and free amino acid release from the active site. Here, we present the X-ray crystal structure of the hexameric leucine aminopeptidase, PfA-M17, alone and in complex with two inhibitors with antimalarial activity. The six active sites of the PfA-M17 hexamer are arranged in a disc-like fashion so that they are orientated inwards to form a central catalytic cavity; flexible loops that sit at each of the six entrances to the catalytic cavern function to regulate substrate access. In stark contrast to PfA-M1, PfA-M17 has a narrow and hydrophobic primary specificity pocket which accounts for its highly restricted substrate specificity. We also explicate the essential roles for the metal-binding centers in these enzymes (two in PfA-M17 and one in PfA-M1) in both substrate and drug binding. Our detailed understanding of the PfA-M1 and PfA-M17 active sites now permits a rational approach in the development of a unique class of two-target and/or combination antimalarial therapy.

• Authors: Sheena McGowan, Christine A Oellig, Woldeamanuel A Birru, Tom T Caradoc-Davies, Colin M Stack, Jonathan Lowther, Tina Skinner-Adams, Artur Mucha, Pawel Kafarski, Jolanta Grembecka, Katharine R Trenholme, Ashley M Buckle, Donald L Gardiner, John P Dalton, James C Whisstock
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 107 Issue 6 Pg. 2449-54 (Feb 09 2010) ISSN: 1091-6490 [Electronic] United States
• PMID: 20133789 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antimalarials
• Metals
• Protozoan Proteins
• Aminopeptidases

Topics

• Aminopeptidases (antagonists & inhibitors, chemistry, metabolism)
• Antimalarials (chemistry, metabolism, pharmacology)
• Catalysis
• Catalytic Domain
• Crystallography, X-Ray
• Drug Design
• Hydrophobic and Hydrophilic Interactions
• Metals (chemistry, metabolism)
• Models, Molecular
• Plasmodium falciparum (drug effects, enzymology, genetics)
• Protein Binding
• Protein Multimerization
• Protein Structure, Quaternary
• Protein Structure, Tertiary
• Protozoan Proteins (antagonists & inhibitors, chemistry, metabolism)
• Substrate Specificity