The M17 leucine aminopeptidase of the malaria parasite Plasmodium falciparum: Importance of active site metal ions in the binding of substrates and inhibitors

Maric, S; Donnelly, SM; Robinson, MW; Skinner-Adams, T; Trenholme, KR; Gardiner, DL; Dalton, JP; Stack, CM; Lowther, J

The M17 leucine aminopeptidase of the malaria parasite Plasmodium falciparum: Importance of active site metal ions in the binding of substrates and inhibitors

Maric, S

Donnelly, SM

Robinson, MW

Skinner-Adams, T Trenholme, KR Gardiner, DL Dalton, JP Stack, CM Lowther, J

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Publication Type:: Journal Article
Citation:: Biochemistry, 2009, 48 (23), pp. 5435 - 5439
Issue Date:: 2009-06-16

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Field	Value	Language
dc.contributor.author	Maric, S https://orcid.org/0000-0001-6547-5427	en_US
dc.contributor.author	Donnelly, SM https://orcid.org/0000-0003-2005-3698	en_US
dc.contributor.author	Robinson, MW https://orcid.org/0000-0001-7191-0034	en_US
dc.contributor.author	Skinner-Adams, T	en_US
dc.contributor.author	Trenholme, KR	en_US
dc.contributor.author	Gardiner, DL	en_US
dc.contributor.author	Dalton, JP	en_US
dc.contributor.author	Stack, CM	en_US
dc.contributor.author	Lowther, J	en_US
dc.date.issued	2009-06-16	en_US
dc.identifier.citation	Biochemistry, 2009, 48 (23), pp. 5435 - 5439	en_US
dc.identifier.issn	0006-2960	en_US
dc.identifier.uri	http://hdl.handle.net/10453/8789
dc.description.abstract	The M17 leucine aminopeptidase of the intraerythrocytic stages of the malaria parasite Plasmodium falciparum (PfLAP) plays a role in releasing amino acids from host hemoglobin that are used for parasite protein synthesis, growth, and development. This enzyme represents a target at which new antimalarials could be designed since metalloaminopeptidase inhibitors prevent the growth of the parasites in vitro and in vivo. A study on the metal ion binding characteristics of recombinant P. falciparum M17 leucine aminopeptidase (rPfLAP) shows that the active site of this exopeptidase contains two metal-binding sites, a readily exchangeable site (site 1) and a tight binding site (site 2). The enzyme retains activity when the metal ion is removed from site 1, while removal of metal ions from both sites results in an inactive apoenzyme that cannot be reactivated by the addition of divalent metal cations. The metal ion at site 1 is readily exchangeable with several divalent metal ions and displays a preference in the order of preference Zn 2+ > Mn 2+ > Co 2+ > Mg 2+. While it is likely that native PfLAP contains a Zn 2+ in site 2, the metal ion located in site 1 may be dependent on the type and concentration of metal ions in the cytosolic compartment of the parasite. Importantly, the type of metal ion present at site 1 influences not only the catalytic efficiency of the enzyme for peptide substrates but also the mode of binding by bestatin, a metal-chelating inhibitor of M17 aminopeptidases with antimalarial activity. © 2009 American Chemical Society.	en_US
dc.relation.ispartof	Biochemistry	en_US
dc.relation.isbasedon	10.1021/bi9003638	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Plasmodium falciparum	en_US
dc.subject.mesh	Metals	en_US
dc.subject.mesh	Leucyl Aminopeptidase	en_US
dc.subject.mesh	Recombinant Proteins	en_US
dc.subject.mesh	Enzyme Inhibitors	en_US
dc.subject.mesh	Binding Sites	en_US
dc.subject.mesh	Catalytic Domain	en_US
dc.subject.mesh	Structure-Activity Relationship	en_US
dc.subject.mesh	Substrate Specificity	en_US
dc.subject.mesh	Kinetics	en_US
dc.title	The M17 leucine aminopeptidase of the malaria parasite Plasmodium falciparum: Importance of active site metal ions in the binding of substrates and inhibitors	en_US
dc.type	Journal Article
utslib.citation.volume	23	en_US
utslib.citation.volume	48	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	0304 Medicinal and Biomolecular Chemistry	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
utslib.for	1101 Medical Biochemistry and Metabolomics	en_US
dc.location.activity	ISI:000266860400044	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
utslib.copyright.status	closed_access
pubs.issue	23	en_US
pubs.publication-status	Published	en_US
pubs.volume	48	en_US

Abstract:

The M17 leucine aminopeptidase of the intraerythrocytic stages of the malaria parasite Plasmodium falciparum (PfLAP) plays a role in releasing amino acids from host hemoglobin that are used for parasite protein synthesis, growth, and development. This enzyme represents a target at which new antimalarials could be designed since metalloaminopeptidase inhibitors prevent the growth of the parasites in vitro and in vivo. A study on the metal ion binding characteristics of recombinant P. falciparum M17 leucine aminopeptidase (rPfLAP) shows that the active site of this exopeptidase contains two metal-binding sites, a readily exchangeable site (site 1) and a tight binding site (site 2). The enzyme retains activity when the metal ion is removed from site 1, while removal of metal ions from both sites results in an inactive apoenzyme that cannot be reactivated by the addition of divalent metal cations. The metal ion at site 1 is readily exchangeable with several divalent metal ions and displays a preference in the order of preference Zn 2+ > Mn 2+ > Co 2+ > Mg 2+. While it is likely that native PfLAP contains a Zn 2+ in site 2, the metal ion located in site 1 may be dependent on the type and concentration of metal ions in the cytosolic compartment of the parasite. Importantly, the type of metal ion present at site 1 influences not only the catalytic efficiency of the enzyme for peptide substrates but also the mode of binding by bestatin, a metal-chelating inhibitor of M17 aminopeptidases with antimalarial activity. © 2009 American Chemical Society.

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http://hdl.handle.net/10453/8789