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
Abstract
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.
- Authors:
-
- Baylor College of Medicine, Houston, TX (United States); Univ. of Kassel, Hesse (Germany)
- London School of Hygiene & Tropical Medicine, London (United Kingdom)
- Univ. of Kassel, Hesse (Germany)
- Emory Univ., Atlanta, GA (United States); Baylor College of Medicine, Houston, TX (United States)
- Baylor College of Medicine, Houston, TX (United States)
- Univ. of Geneva (Switzerland)
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- Wellcome Trust; National Institutes of Health (NIH)
- OSTI Identifier:
- 1234756
- Grant/Contract Number:
- 241481; R01 GM090161; R21 HL111953
- Resource Type:
- Accepted Manuscript
- Journal Name:
- PLoS Pathogens
- Additional Journal Information:
- Journal Volume: 11; Journal Issue: 2; Journal ID: ISSN 1553-7374
- Publisher:
- Public Library of Science
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; crystal structure; parasitic life cycles; malarial parasites; plasmodium; blood; merozoites; hydrogen bonding; red blood cells
Citation Formats
Kim, Jeong Joo, Flueck, Christian, Franz, Eugen, Sanabria-Figueroa, Eduardo, Thompson, Eloise, Lorenz, Robin, Bertinetti, Daniela, Baker, David A., Herberg, Friedrich W., Kim, Choel, and Soldati-Favre, Dominique. 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. United States: N. p., 2015.
Web. doi:10.1371/journal.ppat.1004639.
Kim, Jeong Joo, Flueck, Christian, Franz, Eugen, Sanabria-Figueroa, Eduardo, Thompson, Eloise, Lorenz, Robin, Bertinetti, Daniela, Baker, David A., Herberg, Friedrich W., Kim, Choel, & Soldati-Favre, Dominique. 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. United States. https://doi.org/10.1371/journal.ppat.1004639
Kim, Jeong Joo, Flueck, Christian, Franz, Eugen, Sanabria-Figueroa, Eduardo, Thompson, Eloise, Lorenz, Robin, Bertinetti, Daniela, Baker, David A., Herberg, Friedrich W., Kim, Choel, and Soldati-Favre, Dominique. Tue .
"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". United States. https://doi.org/10.1371/journal.ppat.1004639. https://www.osti.gov/servlets/purl/1234756.
@article{osti_1234756,
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},
author = {Kim, Jeong Joo and Flueck, Christian and Franz, Eugen and Sanabria-Figueroa, Eduardo and Thompson, Eloise and Lorenz, Robin and Bertinetti, Daniela and Baker, David A. and Herberg, Friedrich W. and Kim, Choel and Soldati-Favre, Dominique},
abstractNote = {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.},
doi = {10.1371/journal.ppat.1004639},
journal = {PLoS Pathogens},
number = 2,
volume = 11,
place = {United States},
year = {Tue Feb 03 00:00:00 EST 2015},
month = {Tue Feb 03 00:00:00 EST 2015}
}
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