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

Sheena McGowan, Christine Oellig, Woldeamanuel A Birru, Tom T Caradoc-Davies, Colin M Stack, Jonathan Lowther, Tina S Skinner-Adams, Artur Mucha, Pawel Kafarski, Jolanta Grembecka, Katharine R Trenholme, Ashley Maurice Buckle, Donald L Gardiner, John P Dalton, James Whisstock

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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.
Original languageEnglish
Pages (from-to)2449 - 2454
Number of pages6
JournalProceedings of the National Academy of Sciences
Volume107
Issue number6
DOIs
Publication statusPublished - 2010

Cite this

McGowan, Sheena ; Oellig, Christine ; Birru, Woldeamanuel A ; Caradoc-Davies, Tom T ; Stack, Colin M ; Lowther, Jonathan ; Skinner-Adams, Tina S ; Mucha, Artur ; Kafarski, Pawel ; Grembecka, Jolanta ; Trenholme, Katharine R ; Buckle, Ashley Maurice ; Gardiner, Donald L ; Dalton, John P ; Whisstock, James. / Structure of the Plasmodium falciparum M17 aminopeptidase and significance for the design of drugs targeting the neutral exopeptidases. In: Proceedings of the National Academy of Sciences. 2010 ; Vol. 107, No. 6. pp. 2449 - 2454.
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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.",
author = "Sheena McGowan and Christine Oellig and Birru, {Woldeamanuel A} and Caradoc-Davies, {Tom T} and Stack, {Colin M} and Jonathan Lowther and Skinner-Adams, {Tina S} and Artur Mucha and Pawel Kafarski and Jolanta Grembecka and Trenholme, {Katharine R} and Buckle, {Ashley Maurice} and Gardiner, {Donald L} and Dalton, {John P} and James Whisstock",
year = "2010",
doi = "10.1073/pnas.0911813107",
language = "English",
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McGowan, S, Oellig, C, Birru, WA, Caradoc-Davies, TT, Stack, CM, Lowther, J, Skinner-Adams, TS, Mucha, A, Kafarski, P, Grembecka, J, Trenholme, KR, Buckle, AM, Gardiner, DL, Dalton, JP & Whisstock, J 2010, 'Structure of the Plasmodium falciparum M17 aminopeptidase and significance for the design of drugs targeting the neutral exopeptidases', Proceedings of the National Academy of Sciences, vol. 107, no. 6, pp. 2449 - 2454. https://doi.org/10.1073/pnas.0911813107

Structure of the Plasmodium falciparum M17 aminopeptidase and significance for the design of drugs targeting the neutral exopeptidases. / McGowan, Sheena; Oellig, Christine; Birru, Woldeamanuel A; Caradoc-Davies, Tom T; Stack, Colin M; Lowther, Jonathan; Skinner-Adams, Tina S; Mucha, Artur; Kafarski, Pawel; Grembecka, Jolanta; Trenholme, Katharine R; Buckle, Ashley Maurice; Gardiner, Donald L; Dalton, John P; Whisstock, James.

In: Proceedings of the National Academy of Sciences, Vol. 107, No. 6, 2010, p. 2449 - 2454.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Structure of the Plasmodium falciparum M17 aminopeptidase and significance for the design of drugs targeting the neutral exopeptidases

AU - McGowan, Sheena

AU - Oellig, Christine

AU - Birru, Woldeamanuel A

AU - Caradoc-Davies, Tom T

AU - Stack, Colin M

AU - Lowther, Jonathan

AU - Skinner-Adams, Tina S

AU - Mucha, Artur

AU - Kafarski, Pawel

AU - Grembecka, Jolanta

AU - Trenholme, Katharine R

AU - Buckle, Ashley Maurice

AU - Gardiner, Donald L

AU - Dalton, John P

AU - Whisstock, James

PY - 2010

Y1 - 2010

N2 - 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.

AB - 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.

UR - http://www.pnas.org/content/early/2010/01/11/0911813107.full.pdf+html

U2 - 10.1073/pnas.0911813107

DO - 10.1073/pnas.0911813107

M3 - Article

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EP - 2454

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

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