title: Phosphoproteomics reveals malaria parasite Protein Kinase G as a signalling hub regulating egress and invasion.
abstract: Our understanding of the key phosphorylation-dependent signalling pathways in the human malaria parasite, Plasmodium falciparum, remains rudimentary. Here we address this issue for the essential cGMP-dependent protein kinase, PfPKG. By employing chemical and genetic tools in combination with quantitative global phosphoproteomics, we identify the phosphorylation sites on 69 proteins that are direct or indirect cellular targets for PfPKG. These PfPKG targets include proteins involved in cell signalling, proteolysis, gene regulation, protein export and ion and protein transport, indicating that cGMP/PfPKG acts as a signalling hub that plays a central role in a number of core parasite processes. We also show that PfPKG activity is required for parasite invasion. This correlates with the finding that the calcium-dependent protein kinase, PfCDPK1, is phosphorylated by PfPKG, as are components of the actomyosin complex, providing mechanistic insight into the essential role of PfPKG in parasite egress and invasion.
authors.affiliation: MRC Toxicology Unit, University of Leicester, Hodgkin Building, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: Mahmood M
authors.initials: MM
authors.lastname: Alam
authors.affiliation: MRC Toxicology Unit, University of Leicester, Hodgkin Building, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: Lev
authors.initials: L
authors.lastname: Solyakov
authors.affiliation: PNACL, Core Biotechnology Services, University of Leicester, Hodgkin Building, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: Andrew R
authors.identifier: 0000000251823643
authors.initials: AR
authors.lastname: Bottrill
authors.affiliation: Faculty of Infectious and Tropical Diseases, London School of Hygiene &Tropical Medicine, Keppel Street, London WC1E 7HT, UK.
authors.forename: Christian
authors.initials: C
authors.lastname: Flueck
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi 110 067, India.
authors.forename: Faiza A
authors.initials: FA
authors.lastname: Siddiqui
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi 110 067, India.
authors.forename: Shailja
authors.initials: S
authors.lastname: Singh
authors.affiliation: PNACL, Core Biotechnology Services, University of Leicester, Hodgkin Building, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: Sharad
authors.initials: S
authors.lastname: Mistry
authors.affiliation: Bioinformatics and Biostatistics Support Hub (B/BASH), Core Biotechnology Services, Maurice Shock Building, University of Leicester, University Road, Leicester LE1 9HN, UK.
authors.forename: Maria
authors.initials: M
authors.lastname: Viskaduraki
authors.affiliation: Bioinformatics and Biostatistics Support Hub (B/BASH), Core Biotechnology Services, Maurice Shock Building, University of Leicester, University Road, Leicester LE1 9HN, UK.
authors.forename: Kate
authors.initials: K
authors.lastname: Lee
authors.affiliation: Faculty of Infectious and Tropical Diseases, London School of Hygiene &Tropical Medicine, Keppel Street, London WC1E 7HT, UK.
authors.forename: Christine S
authors.initials: CS
authors.lastname: Hopp
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi 110 067, India.
authors.forename: Chetan E
authors.initials: CE
authors.lastname: Chitnis
authors.affiliation: Department of Microbiology, Monash University, Building 76, Clayton, Victoria 3800, Australia.
authors.forename: Christian
authors.initials: C
authors.lastname: Doerig
authors.affiliation: Division of Parasitology, MRC National Institute for Medical Research, Ridgeway, Mill Hill, London NW7 1AA, UK.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Moon
authors.affiliation: Division of Parasitology, MRC National Institute for Medical Research, Ridgeway, Mill Hill, London NW7 1AA, UK.
authors.forename: Judith L
authors.initials: JL
authors.lastname: Green
authors.affiliation: Division of Parasitology, MRC National Institute for Medical Research, Ridgeway, Mill Hill, London NW7 1AA, UK.
authors.forename: Anthony A
authors.initials: AA
authors.lastname: Holder
authors.affiliation: Faculty of Infectious and Tropical Diseases, London School of Hygiene &Tropical Medicine, Keppel Street, London WC1E 7HT, UK.
authors.forename: David A
authors.initials: DA
authors.lastname: Baker
authors.affiliation: MRC Toxicology Unit, University of Leicester, Hodgkin Building, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Tobin
mesh_terms: D020013:Calcium Signaling / Q000502:physiology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D004912:Erythrocytes / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D052939:Schizonts / Q000502:physiology
chemical_list: D010750:Phosphoproteins; D015800:Protozoan Proteins; D017869:Cyclic GMP-Dependent Protein Kinases