Fingerprinting the substrate specificity of M1 and M17 aminopeptidases of human malaria, Plasmodium falciparum.

Abstract

BACKGROUND:

Plasmodium falciparum, the causative agent of human malaria, expresses two aminopeptidases, PfM1AAP and PfM17LAP, critical to generating a free amino acid pool used by the intraerythrocytic stage of the parasite for proteins synthesis, growth and development. These exopeptidases are potential targets for the development of a new class of anti-malaria drugs.

METHODOLOGY/PRINCIPAL FINDINGS:

To define the substrate specificity of recombinant forms of these two malaria aminopeptidases we used a new library consisting of 61 fluorogenic substrates derived both from natural and unnatural amino acids. We obtained a detailed substrate fingerprint for recombinant forms of the enzymes revealing that PfM1AAP exhibits a very broad substrate tolerance, capable of efficiently hydrolyzing neutral and basic amino acids, while PfM17LAP has narrower substrate specificity and preferentially cleaves bulky, hydrophobic amino acids. The substrate library was also exploited to profile the activity of the native aminopeptidases in soluble cell lysates of P. falciparum malaria.

CONCLUSIONS/SIGNIFICANCE:

This data showed that PfM1AAP and PfM17LAP are responsible for majority of the aminopeptidase activity in these extracts. These studies provide specific substrate and mechanistic information important for understanding the function of these aminopeptidases and could be exploited in the design of new inhibitors to specifically target these for anti-malaria treatment.

• Authors: Marcin Poreba, Sheena McGowan, Tina S Skinner-Adams, Katharine R Trenholme, Donald L Gardiner, James C Whisstock, Joyce To, Guy S Salvesen, John P Dalton, Marcin Drag
• Journal: PloS one (PLoS One) Vol. 7 Issue 2 Pg. e31938 ( 2012) ISSN: 1932-6203 [Electronic] United States
• PMID: 22359643 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Amino Acids
• Small Molecule Libraries
• Aminopeptidases

Topics

• Amino Acids (chemistry)
• Aminopeptidases (chemistry)
• Drug Design
• Humans
• Malaria, Falciparum (drug therapy, enzymology)
• Molecular Targeted Therapy
• Plasmodium falciparum (enzymology)
• Small Molecule Libraries
• Substrate Specificity