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Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum

Red blood cell invasion by the malaria parasite Plasmodium falciparum relies on a complex protein network that uses low and high affinity receptor–ligand interactions. Signal transduction through the action of specific kinases is a control mechanism for the orchestration of this process. In the pres...

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Autores principales: Engelberg, Klemens, Paul, Aditya S., Prinz, Boris, Kono, Maya, Ching, Wilhelm, Heincke, Dorothee, Dobner, Thomas, Spielmann, Tobias, Duraisingh, Manoj T., Gilberger, Tim-Wolf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3671792/
https://www.ncbi.nlm.nih.gov/pubmed/23544851
http://dx.doi.org/10.1042/BJ20121694
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author Engelberg, Klemens
Paul, Aditya S.
Prinz, Boris
Kono, Maya
Ching, Wilhelm
Heincke, Dorothee
Dobner, Thomas
Spielmann, Tobias
Duraisingh, Manoj T.
Gilberger, Tim-Wolf
author_facet Engelberg, Klemens
Paul, Aditya S.
Prinz, Boris
Kono, Maya
Ching, Wilhelm
Heincke, Dorothee
Dobner, Thomas
Spielmann, Tobias
Duraisingh, Manoj T.
Gilberger, Tim-Wolf
author_sort Engelberg, Klemens
collection PubMed
description Red blood cell invasion by the malaria parasite Plasmodium falciparum relies on a complex protein network that uses low and high affinity receptor–ligand interactions. Signal transduction through the action of specific kinases is a control mechanism for the orchestration of this process. In the present study we report on the phosphorylation of the CPD (cytoplasmic domain) of P. falciparum Rh2b (reticulocyte homologue protein 2b). First, we identified Ser(3233) as the sole phospho-acceptor site in the CPD for in vitro phosphorylation by parasite extract. We provide several lines of evidence that this phosphorylation is mediated by PfCK2 (P. falciparum casein kinase 2): phosphorylation is cAMP independent, utilizes ATP as well as GTP as phosphate donors, is inhibited by heparin and tetrabromocinnamic acid, and is mediated by purified PfCK2. We raised a phospho-specific antibody and showed that Ser(3233) phosphorylation occurs in the parasite prior to host cell egress. We analysed the spatiotemporal aspects of this phosphorylation using immunoprecipitated endogenous Rh2b and minigenes expressing the CPD either at the plasma or rhoptry membrane. Phosphorylation of Rh2b is not spatially restricted to either the plasma or rhoptry membrane and most probably occurs before Rh2b is translocated from the rhoptry neck to the plasma membrane.
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spelling pubmed-36717922013-06-07 Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum Engelberg, Klemens Paul, Aditya S. Prinz, Boris Kono, Maya Ching, Wilhelm Heincke, Dorothee Dobner, Thomas Spielmann, Tobias Duraisingh, Manoj T. Gilberger, Tim-Wolf Biochem J Research Article Red blood cell invasion by the malaria parasite Plasmodium falciparum relies on a complex protein network that uses low and high affinity receptor–ligand interactions. Signal transduction through the action of specific kinases is a control mechanism for the orchestration of this process. In the present study we report on the phosphorylation of the CPD (cytoplasmic domain) of P. falciparum Rh2b (reticulocyte homologue protein 2b). First, we identified Ser(3233) as the sole phospho-acceptor site in the CPD for in vitro phosphorylation by parasite extract. We provide several lines of evidence that this phosphorylation is mediated by PfCK2 (P. falciparum casein kinase 2): phosphorylation is cAMP independent, utilizes ATP as well as GTP as phosphate donors, is inhibited by heparin and tetrabromocinnamic acid, and is mediated by purified PfCK2. We raised a phospho-specific antibody and showed that Ser(3233) phosphorylation occurs in the parasite prior to host cell egress. We analysed the spatiotemporal aspects of this phosphorylation using immunoprecipitated endogenous Rh2b and minigenes expressing the CPD either at the plasma or rhoptry membrane. Phosphorylation of Rh2b is not spatially restricted to either the plasma or rhoptry membrane and most probably occurs before Rh2b is translocated from the rhoptry neck to the plasma membrane. Portland Press Ltd. 2013-05-31 2013-06-15 /pmc/articles/PMC3671792/ /pubmed/23544851 http://dx.doi.org/10.1042/BJ20121694 Text en © 2013 The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Licence (CC-BY)(http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Engelberg, Klemens
Paul, Aditya S.
Prinz, Boris
Kono, Maya
Ching, Wilhelm
Heincke, Dorothee
Dobner, Thomas
Spielmann, Tobias
Duraisingh, Manoj T.
Gilberger, Tim-Wolf
Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum
title Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum
title_full Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum
title_fullStr Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum
title_full_unstemmed Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum
title_short Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum
title_sort specific phosphorylation of the pfrh2b invasion ligand of plasmodium falciparum
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3671792/
https://www.ncbi.nlm.nih.gov/pubmed/23544851
http://dx.doi.org/10.1042/BJ20121694
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