Characterization of the Plasmodium falciparum M17 leucyl aminopeptidase: A protease involved in amino acid regulation with potential for antimalarial drug development

Stack, CM; Lowther, J; Cunningham, E; Donnelly, S; Gardiner, DL; Trenholme, KR; Skinner-Adams, TS; Teuscher, F; Grembecka, J; Mucha, A; Kafarski, P; Lua, L; Bell, A; Dalton, JP

Characterization of the Plasmodium falciparum M17 leucyl aminopeptidase: A protease involved in amino acid regulation with potential for antimalarial drug development

Stack, CM Lowther, J Cunningham, E Donnelly, S

Gardiner, DL Trenholme, KR Skinner-Adams, TS Teuscher, F Grembecka, J Mucha, A Kafarski, P Lua, L Bell, A Dalton, JP

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Publication Type:: Journal Article
Citation:: Journal of Biological Chemistry, 2007, 282 (3), pp. 2069 - 2080
Issue Date:: 2007-01-19

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Field	Value	Language
dc.contributor.author	Stack, CM	en_US
dc.contributor.author	Lowther, J	en_US
dc.contributor.author	Cunningham, E	en_US
dc.contributor.author	Donnelly, S https://orcid.org/0000-0003-2005-3698	en_US
dc.contributor.author	Gardiner, DL	en_US
dc.contributor.author	Trenholme, KR	en_US
dc.contributor.author	Skinner-Adams, TS	en_US
dc.contributor.author	Teuscher, F	en_US
dc.contributor.author	Grembecka, J	en_US
dc.contributor.author	Mucha, A	en_US
dc.contributor.author	Kafarski, P	en_US
dc.contributor.author	Lua, L	en_US
dc.contributor.author	Bell, A	en_US
dc.contributor.author	Dalton, JP	en_US
dc.date.issued	2007-01-19	en_US
dc.identifier.citation	Journal of Biological Chemistry, 2007, 282 (3), pp. 2069 - 2080	en_US
dc.identifier.issn	0021-9258	en_US
dc.identifier.uri	http://hdl.handle.net/10453/3558
dc.description.abstract	Amino acids generated from the catabolism of hemoglobin by intra-erythrocytic malaria parasites are not only essential for protein synthesis but also function in maintaining an osmotically stable environment, and creating a gradient by which amino acids that are rare or not present in hemoglobin are drawn into the parasite from host serum. We have proposed that a Plasmodium falciparum M17 leucyl aminopeptidase (PfLAP) generates and regulates the internal pool of free amino acids and therefore represents a target for novel antimalarial drugs. This enzyme has been expressed in insect cells as a functional 320-kDa homo-hexamer that is optimally active at neutral or alkaline pH, is dependent on metal ions for activity, and exhibits a substrate preference for N-terminally exposed hydrophobic amino acids, particularly leucine. PfLAP is produced by all stages in the intra-erythrocytic developmental cycle of malaria but was most highly expressed by trophozoites, a stage at which hemoglobin degradation and parasite protein synthesis are elevated. The enzyme was located by immunohistochemical methods and by transfecting malaria cells with a PfLAP-green fluorescent protein construct, to the cytosolic compartment of the cell at all developmental stages, including segregated merozoites. Amino acid dipeptide analogs, such as bestatin and its derivatives, are potent inhibitors of the protease and also block the growth of P. falciparum malaria parasites in culture. This study provides a biochemical basis for the antimalarial activity of aminopeptidase inhibitors. Availability of functionally active recombinant PfLAP, coupled with a simple enzymatic readout, will aid medicinal chemistry and/or high throughput approaches for the future design/discovery of new antimalarial drugs. © 2007 by The American Society for Biochemistry and Molecular Biology, Inc.	en_US
dc.relation.ispartof	Journal of Biological Chemistry	en_US
dc.relation.isbasedon	10.1074/jbc.M609251200	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Cytosol	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Plasmodium falciparum	en_US
dc.subject.mesh	Ions	en_US
dc.subject.mesh	Leucyl Aminopeptidase	en_US
dc.subject.mesh	Amino Acids	en_US
dc.subject.mesh	Leucine	en_US
dc.subject.mesh	Recombinant Proteins	en_US
dc.subject.mesh	Antimalarials	en_US
dc.subject.mesh	Immunohistochemistry	en_US
dc.subject.mesh	Phylogeny	en_US
dc.subject.mesh	Kinetics	en_US
dc.subject.mesh	Plasmids	en_US
dc.subject.mesh	Hydrogen-Ion Concentration	en_US
dc.subject.mesh	Trophozoites	en_US
dc.subject.mesh	Merozoites	en_US
dc.title	Characterization of the Plasmodium falciparum M17 leucyl aminopeptidase: A protease involved in amino acid regulation with potential for antimalarial drug development	en_US
dc.type	Journal Article
utslib.citation.volume	3	en_US
utslib.citation.volume	282	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	03 Chemical Sciences	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	11 Medical and Health Sciences	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	3	en_US
pubs.publication-status	Published	en_US
pubs.volume	282	en_US

Abstract:

Amino acids generated from the catabolism of hemoglobin by intra-erythrocytic malaria parasites are not only essential for protein synthesis but also function in maintaining an osmotically stable environment, and creating a gradient by which amino acids that are rare or not present in hemoglobin are drawn into the parasite from host serum. We have proposed that a Plasmodium falciparum M17 leucyl aminopeptidase (PfLAP) generates and regulates the internal pool of free amino acids and therefore represents a target for novel antimalarial drugs. This enzyme has been expressed in insect cells as a functional 320-kDa homo-hexamer that is optimally active at neutral or alkaline pH, is dependent on metal ions for activity, and exhibits a substrate preference for N-terminally exposed hydrophobic amino acids, particularly leucine. PfLAP is produced by all stages in the intra-erythrocytic developmental cycle of malaria but was most highly expressed by trophozoites, a stage at which hemoglobin degradation and parasite protein synthesis are elevated. The enzyme was located by immunohistochemical methods and by transfecting malaria cells with a PfLAP-green fluorescent protein construct, to the cytosolic compartment of the cell at all developmental stages, including segregated merozoites. Amino acid dipeptide analogs, such as bestatin and its derivatives, are potent inhibitors of the protease and also block the growth of P. falciparum malaria parasites in culture. This study provides a biochemical basis for the antimalarial activity of aminopeptidase inhibitors. Availability of functionally active recombinant PfLAP, coupled with a simple enzymatic readout, will aid medicinal chemistry and/or high throughput approaches for the future design/discovery of new antimalarial drugs. © 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

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http://hdl.handle.net/10453/3558