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 language | English |
|---|---|
| Pages (from-to) | 2112-2123 |
| Number of pages | 12 |
| Journal | Journal of Proteome Research |
| Volume | 17 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 1 Jun 2018 |
Keywords
- intraerythrocytic cycle
- isobaric tags
- kinase-substrate pairs
- malaria
- PfPK7
- phosphoproteomics
- phosphorylation
- Plasmodium falciparum
- TMT
Cite this
}
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 journal › Article › Research › peer-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
UR - http://www.scopus.com/inward/record.url?scp=85046469910&partnerID=8YFLogxK
U2 - 10.1021/acs.jproteome.8b00062
DO - 10.1021/acs.jproteome.8b00062
M3 - Article
VL - 17
SP - 2112
EP - 2123
JO - Journal of Proteome Research
JF - Journal of Proteome Research
SN - 1535-3893
IS - 6
ER -