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

Poreba, M; McGowan, S; Skinner-Adams, TS; Trenholme, KR; Gardiner, DL; Whisstock, JC; To, J; Salvesen, GS; Dalton, JP; Drag, M

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

Poreba, M McGowan, S Skinner-Adams, TS Trenholme, KR Gardiner, DL Whisstock, JC To, J

Salvesen, GS Dalton, JP Drag, M

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Publication Type:: Journal Article
Citation:: PLoS ONE, 2012, 7 (2)
Issue Date:: 2012-02-16

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Field	Value	Language
dc.contributor.author	Poreba, M	en_US
dc.contributor.author	McGowan, S	en_US
dc.contributor.author	Skinner-Adams, TS	en_US
dc.contributor.author	Trenholme, KR	en_US
dc.contributor.author	Gardiner, DL	en_US
dc.contributor.author	Whisstock, JC	en_US
dc.contributor.author	To, J https://orcid.org/0000-0003-3482-4369	en_US
dc.contributor.author	Salvesen, GS	en_US
dc.contributor.author	Dalton, JP	en_US
dc.contributor.author	Drag, M	en_US
dc.date.available	2012-01-18	en_US
dc.date.issued	2012-02-16	en_US
dc.identifier.citation	PLoS ONE, 2012, 7 (2)	en_US
dc.identifier.uri	http://hdl.handle.net/10453/114794
dc.description.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. © 2012 Poreba et al.	en_US
dc.relation.ispartof	PLoS ONE	en_US
dc.relation.isbasedon	10.1371/journal.pone.0031938	en_US
dc.subject.classification	General Science & Technology	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Plasmodium falciparum	en_US
dc.subject.mesh	Malaria, Falciparum	en_US
dc.subject.mesh	Aminopeptidases	en_US
dc.subject.mesh	Amino Acids	en_US
dc.subject.mesh	Substrate Specificity	en_US
dc.subject.mesh	Drug Design	en_US
dc.subject.mesh	Small Molecule Libraries	en_US
dc.subject.mesh	Molecular Targeted Therapy	en_US
dc.title	Fingerprinting the substrate specificity of M1 and M17 aminopeptidases of human malaria, Plasmodium falciparum	en_US
dc.type	Journal Article
utslib.citation.volume	2	en_US
utslib.citation.volume	7	en_US
utslib.for	0605 Microbiology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	open_access
pubs.issue	2	en_US
pubs.publication-status	Published	en_US
pubs.volume	7	en_US

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. © 2012 Poreba et al.

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http://hdl.handle.net/10453/114794