Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress.

The Plasmodium falciparum cGMP-dependent protein kinase (PfPKG) is a key regulator across the malaria parasite life cycle. Little is known about PfPKG's activation mechanism. Here we report that the carboxyl cyclic nucleotide binding domain functions as a "gatekeeper" for activation b...

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Main Authors: Jeong Joo Kim, Christian Flueck, Eugen Franz, Eduardo Sanabria-Figueroa, Eloise Thompson, Robin Lorenz, Daniela Bertinetti, David A Baker, Friedrich W Herberg, Choel Kim
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2015-02-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1004639
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author Jeong Joo Kim
Christian Flueck
Eugen Franz
Eduardo Sanabria-Figueroa
Eloise Thompson
Robin Lorenz
Daniela Bertinetti
David A Baker
Friedrich W Herberg
Choel Kim
author_facet Jeong Joo Kim
Christian Flueck
Eugen Franz
Eduardo Sanabria-Figueroa
Eloise Thompson
Robin Lorenz
Daniela Bertinetti
David A Baker
Friedrich W Herberg
Choel Kim
author_sort Jeong Joo Kim
collection DOAJ
description The Plasmodium falciparum cGMP-dependent protein kinase (PfPKG) is a key regulator across the malaria parasite life cycle. Little is known about PfPKG's activation mechanism. Here we report that the carboxyl cyclic nucleotide binding domain functions as a "gatekeeper" for activation by providing the highest cGMP affinity and selectivity. To understand the mechanism, we have solved its crystal structures with and without cGMP at 2.0 and 1.9 Å, respectively. These structures revealed a PfPKG-specific capping triad that forms upon cGMP binding, and disrupting the triad reduces kinase activity by 90%. Furthermore, mutating these residues in the parasite prevents blood stage merozoite egress, confirming the essential nature of the triad in the parasite. We propose a mechanism of activation where cGMP binding allosterically triggers the conformational change at the αC-helix, which bridges the regulatory and catalytic domains, causing the capping triad to form and stabilize the active conformation.
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institution Kabale University
issn 1553-7366
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language English
publishDate 2015-02-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS Pathogens
spelling doaj-art-cdf948dfee564831ba47e158adaa23ca2025-08-20T03:46:13ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742015-02-01112e100463910.1371/journal.ppat.1004639Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress.Jeong Joo KimChristian FlueckEugen FranzEduardo Sanabria-FigueroaEloise ThompsonRobin LorenzDaniela BertinettiDavid A BakerFriedrich W HerbergChoel KimThe Plasmodium falciparum cGMP-dependent protein kinase (PfPKG) is a key regulator across the malaria parasite life cycle. Little is known about PfPKG's activation mechanism. Here we report that the carboxyl cyclic nucleotide binding domain functions as a "gatekeeper" for activation by providing the highest cGMP affinity and selectivity. To understand the mechanism, we have solved its crystal structures with and without cGMP at 2.0 and 1.9 Å, respectively. These structures revealed a PfPKG-specific capping triad that forms upon cGMP binding, and disrupting the triad reduces kinase activity by 90%. Furthermore, mutating these residues in the parasite prevents blood stage merozoite egress, confirming the essential nature of the triad in the parasite. We propose a mechanism of activation where cGMP binding allosterically triggers the conformational change at the αC-helix, which bridges the regulatory and catalytic domains, causing the capping triad to form and stabilize the active conformation.https://doi.org/10.1371/journal.ppat.1004639
spellingShingle Jeong Joo Kim
Christian Flueck
Eugen Franz
Eduardo Sanabria-Figueroa
Eloise Thompson
Robin Lorenz
Daniela Bertinetti
David A Baker
Friedrich W Herberg
Choel Kim
Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress.
PLoS Pathogens
title Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress.
title_full Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress.
title_fullStr Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress.
title_full_unstemmed Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress.
title_short Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress.
title_sort crystal structures of the carboxyl cgmp binding domain of the plasmodium falciparum cgmp dependent protein kinase reveal a novel capping triad crucial for merozoite egress
url https://doi.org/10.1371/journal.ppat.1004639
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