Characterization of Plasmodium falciparum Atypical Kinase PfPK7- Dependent Phosphoproteome

Brittany N. Pease, Edward L. Huttlin, Mark P. Jedrychowski, Dominique Dorin-Semblat, Daniela Sebastiani, Daniel T. Segarra, Bracken F. Roberts, Ratna Chakrabarti, Christian Doerig, Steven P. Gygi, Debopam Chakrabarti

Research output: Contribution to journalArticleResearchpeer-review

Abstract

PfPK7 is an "orphan" kinase displaying regions of homology to multiple protein kinase families. PfPK7 functions in regulating parasite proliferation/development as evident from the phenotype analysis of knockout parasites. Despite this regulatory role, the functions of PfPK7 in signaling pathways are not known. To better understand PfPK7-regulated phosphorylation events, we performed isobaric tag-based quantitative comparative phosphoproteomics of the schizont and segmenter stages from wild-type and pfpk7- parasite lines. This analysis identified 3,875 phosphorylation sites on 1,047 proteins. Among these phosphorylation events, 146 proteins with 239 phosphorylation sites displayed reduction in phosphorylation in the absence of PfPK7. Further analysis of the phosphopeptides revealed three motifs whose phosphorylation was down regulated in the pfpk7- cell line in both schizonts and segmenters. Decreased phosphorylation following loss of PfPK7 indicates that these proteins may function as direct substrates of PfPK7. We demonstrated that PfPK7 is active toward three of these potential novel substrates; however, PfPK7 did not phosphorylate many of the other proteins, suggesting that decreased phosphorylation in these proteins is an indirect effect. Our phosphoproteomics analysis is the first study to identify direct substrates of PfPK7 and reveals potential downstream or compensatory signaling pathways.

Original languageEnglish
Pages (from-to)2112-2123
Number of pages12
JournalJournal of Proteome Research
Volume17
Issue number6
DOIs
Publication statusPublished - 1 Jun 2018

Keywords

  • intraerythrocytic cycle
  • isobaric tags
  • kinase-substrate pairs
  • malaria
  • PfPK7
  • phosphoproteomics
  • phosphorylation
  • Plasmodium falciparum
  • TMT

Cite this

Pease, B. N., Huttlin, E. L., Jedrychowski, M. P., Dorin-Semblat, D., Sebastiani, D., Segarra, D. T., ... Chakrabarti, D. (2018). Characterization of Plasmodium falciparum Atypical Kinase PfPK7- Dependent Phosphoproteome. Journal of Proteome Research, 17(6), 2112-2123. https://doi.org/10.1021/acs.jproteome.8b00062
Pease, Brittany N. ; Huttlin, Edward L. ; Jedrychowski, Mark P. ; Dorin-Semblat, Dominique ; Sebastiani, Daniela ; Segarra, Daniel T. ; Roberts, Bracken F. ; Chakrabarti, Ratna ; Doerig, Christian ; Gygi, Steven P. ; Chakrabarti, Debopam. / Characterization of Plasmodium falciparum Atypical Kinase PfPK7- Dependent Phosphoproteome. In: Journal of Proteome Research. 2018 ; Vol. 17, No. 6. pp. 2112-2123.
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title = "Characterization of Plasmodium falciparum Atypical Kinase PfPK7- Dependent Phosphoproteome",
abstract = "PfPK7 is an {"}orphan{"} kinase displaying regions of homology to multiple protein kinase families. PfPK7 functions in regulating parasite proliferation/development as evident from the phenotype analysis of knockout parasites. Despite this regulatory role, the functions of PfPK7 in signaling pathways are not known. To better understand PfPK7-regulated phosphorylation events, we performed isobaric tag-based quantitative comparative phosphoproteomics of the schizont and segmenter stages from wild-type and pfpk7- parasite lines. This analysis identified 3,875 phosphorylation sites on 1,047 proteins. Among these phosphorylation events, 146 proteins with 239 phosphorylation sites displayed reduction in phosphorylation in the absence of PfPK7. Further analysis of the phosphopeptides revealed three motifs whose phosphorylation was down regulated in the pfpk7- cell line in both schizonts and segmenters. Decreased phosphorylation following loss of PfPK7 indicates that these proteins may function as direct substrates of PfPK7. We demonstrated that PfPK7 is active toward three of these potential novel substrates; however, PfPK7 did not phosphorylate many of the other proteins, suggesting that decreased phosphorylation in these proteins is an indirect effect. Our phosphoproteomics analysis is the first study to identify direct substrates of PfPK7 and reveals potential downstream or compensatory signaling pathways.",
keywords = "intraerythrocytic cycle, isobaric tags, kinase-substrate pairs, malaria, PfPK7, phosphoproteomics, phosphorylation, Plasmodium falciparum, TMT",
author = "Pease, {Brittany N.} and Huttlin, {Edward L.} and Jedrychowski, {Mark P.} and Dominique Dorin-Semblat and Daniela Sebastiani and Segarra, {Daniel T.} and Roberts, {Bracken F.} and Ratna Chakrabarti and Christian Doerig and Gygi, {Steven P.} and Debopam Chakrabarti",
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Pease, BN, Huttlin, EL, Jedrychowski, MP, Dorin-Semblat, D, Sebastiani, D, Segarra, DT, Roberts, BF, Chakrabarti, R, Doerig, C, Gygi, SP & Chakrabarti, D 2018, 'Characterization of Plasmodium falciparum Atypical Kinase PfPK7- Dependent Phosphoproteome', Journal of Proteome Research, vol. 17, no. 6, pp. 2112-2123. https://doi.org/10.1021/acs.jproteome.8b00062

Characterization of Plasmodium falciparum Atypical Kinase PfPK7- Dependent Phosphoproteome. / Pease, Brittany N.; Huttlin, Edward L.; Jedrychowski, Mark P.; Dorin-Semblat, Dominique; Sebastiani, Daniela; Segarra, Daniel T.; Roberts, Bracken F.; Chakrabarti, Ratna; Doerig, Christian; Gygi, Steven P.; Chakrabarti, Debopam.

In: Journal of Proteome Research, Vol. 17, No. 6, 01.06.2018, p. 2112-2123.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Characterization of Plasmodium falciparum Atypical Kinase PfPK7- Dependent Phosphoproteome

AU - Pease, Brittany N.

AU - Huttlin, Edward L.

AU - Jedrychowski, Mark P.

AU - Dorin-Semblat, Dominique

AU - Sebastiani, Daniela

AU - Segarra, Daniel T.

AU - Roberts, Bracken F.

AU - Chakrabarti, Ratna

AU - Doerig, Christian

AU - Gygi, Steven P.

AU - Chakrabarti, Debopam

PY - 2018/6/1

Y1 - 2018/6/1

N2 - PfPK7 is an "orphan" kinase displaying regions of homology to multiple protein kinase families. PfPK7 functions in regulating parasite proliferation/development as evident from the phenotype analysis of knockout parasites. Despite this regulatory role, the functions of PfPK7 in signaling pathways are not known. To better understand PfPK7-regulated phosphorylation events, we performed isobaric tag-based quantitative comparative phosphoproteomics of the schizont and segmenter stages from wild-type and pfpk7- parasite lines. This analysis identified 3,875 phosphorylation sites on 1,047 proteins. Among these phosphorylation events, 146 proteins with 239 phosphorylation sites displayed reduction in phosphorylation in the absence of PfPK7. Further analysis of the phosphopeptides revealed three motifs whose phosphorylation was down regulated in the pfpk7- cell line in both schizonts and segmenters. Decreased phosphorylation following loss of PfPK7 indicates that these proteins may function as direct substrates of PfPK7. We demonstrated that PfPK7 is active toward three of these potential novel substrates; however, PfPK7 did not phosphorylate many of the other proteins, suggesting that decreased phosphorylation in these proteins is an indirect effect. Our phosphoproteomics analysis is the first study to identify direct substrates of PfPK7 and reveals potential downstream or compensatory signaling pathways.

AB - PfPK7 is an "orphan" kinase displaying regions of homology to multiple protein kinase families. PfPK7 functions in regulating parasite proliferation/development as evident from the phenotype analysis of knockout parasites. Despite this regulatory role, the functions of PfPK7 in signaling pathways are not known. To better understand PfPK7-regulated phosphorylation events, we performed isobaric tag-based quantitative comparative phosphoproteomics of the schizont and segmenter stages from wild-type and pfpk7- parasite lines. This analysis identified 3,875 phosphorylation sites on 1,047 proteins. Among these phosphorylation events, 146 proteins with 239 phosphorylation sites displayed reduction in phosphorylation in the absence of PfPK7. Further analysis of the phosphopeptides revealed three motifs whose phosphorylation was down regulated in the pfpk7- cell line in both schizonts and segmenters. Decreased phosphorylation following loss of PfPK7 indicates that these proteins may function as direct substrates of PfPK7. We demonstrated that PfPK7 is active toward three of these potential novel substrates; however, PfPK7 did not phosphorylate many of the other proteins, suggesting that decreased phosphorylation in these proteins is an indirect effect. Our phosphoproteomics analysis is the first study to identify direct substrates of PfPK7 and reveals potential downstream or compensatory signaling pathways.

KW - intraerythrocytic cycle

KW - isobaric tags

KW - kinase-substrate pairs

KW - malaria

KW - PfPK7

KW - phosphoproteomics

KW - phosphorylation

KW - Plasmodium falciparum

KW - TMT

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U2 - 10.1021/acs.jproteome.8b00062

DO - 10.1021/acs.jproteome.8b00062

M3 - Article

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EP - 2123

JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

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Pease BN, Huttlin EL, Jedrychowski MP, Dorin-Semblat D, Sebastiani D, Segarra DT et al. Characterization of Plasmodium falciparum Atypical Kinase PfPK7- Dependent Phosphoproteome. Journal of Proteome Research. 2018 Jun 1;17(6):2112-2123. https://doi.org/10.1021/acs.jproteome.8b00062