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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

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:
 [1];  [2];  [3];  [4];  [2];  [3];  [3];  [2];  [3];  [5];  [6]
  1. Baylor College of Medicine, Houston, TX (United States); Univ. of Kassel, Hesse (Germany)
  2. London School of Hygiene & Tropical Medicine, London (United Kingdom)
  3. Univ. of Kassel, Hesse (Germany)
  4. Emory Univ., Atlanta, GA (United States); Baylor College of Medicine, Houston, TX (United States)
  5. Baylor College of Medicine, Houston, TX (United States)
  6. Univ. of Geneva (Switzerland)
Publication Date:
Research Org.:
Argonne National Lab. (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. 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, 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. doi: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 = {2015},
month = {2}
}

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    Works referencing / citing this record:

    Structures of the cGMP-dependent protein kinase in malaria parasites reveal a unique structural relay mechanism for activation
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    • El Bakkouri, Majida; Kouidmi, Imène; Wernimont, Amy K.
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