Dataset Information

Phosphoproteomic analysis of Protein Phosphatase PfPPM2/PP2C mutant of Plasmodium falciparum 3D7

ABSTRACT: PfPPM2, a protein phosphatase, plays a key role in regulating both the asexual and sexual development of the human malaria parasite P. falciparum. To understand the mechanisms by which PfPPM2 influences this process, it was essential to identify its targets or substrates within the parasite. To achieve this, we conducted a comparative phosphoproteomic analysis between PfPPM2 depletion mutants and wild-type lines.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Plasmodium Falciparum (isolate 3d7)

SUBMITTER: Keshava Prasad T. S.

LAB HEAD: Prof. Dr. Keshava Prasad T. S.

PROVIDER: PXD055390 | Pride | 2025-04-08

REPOSITORIES: Pride

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Source:

			Action	DRS
	PP2C_Control_line_Phospho.msf	Msf
	PP2C_Control_line_TP.msf	Msf
	PP2C_Mutant_Phospho_F10_R1.raw	Raw
	PP2C_Mutant_Phospho_F10_R2.raw	Raw
	PP2C_Mutant_Phospho_F10_R3.raw	Raw

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Project description:During intra-erythrocytic development, late asexually replicating Plasmodium falciparum parasites sequester from peripheral circulation. This facilitates chronic infection and is linked to severe disease and organ-specific pathology including cerebral and placental malaria. Immature gametocytes M-bM-^@M-^S sexual stage precursor cells M-bM-^@M-^S likewise disappear from circulation. Recent work has demonstrated that these sexual stage parasites are located in the hematopoietic system of the bone marrow before mature gametocytes are released into the blood stream to facilitate mosquito transmission. However, as sequestration occurs only in vivo and not during in vitro culture, the mechanisms by which it is regulated and enacted (particularly by the gametocyte stage) remain poorly understood. We generated the most comprehensive P. falciparum functional gene network to date by integrating global transcriptional data from a large set of asexual and sexual in vitro samples, patient-derived in vivo samples, and a new set of in vitro samples profiling sexual commitment. We defined more than 250 functional modules (clusters) of genes that are co-expressed primarily during the intra-erythrocytic parasite cycle, including 35 during sexual commitment and gametocyte development. Comparing the in vivo and in vitro datasets allowed us, for the first time, to map the time point of asexual parasite sequestration in patients to 22 hours post invasion, confirming previous in vitro observations on the dynamics of host cell modification and cytoadherence. Moreover, we were able to define the properties of gametocyte sequestration, demonstrating the presence of two circulating gametocyte populations: gametocyte rings between 0 and ~30 hours post invasion and mature gametocytes after around 7 days post invasion. We used 164/TdTom, a transgenic parasite line expressing a red fluorescent protein reporter under a gametocyte-specific promoter to generate schizont samples. Schizonts were subsequently isolated from both the fluorescent and non-fluorescent population by FACS and prepared for microarray analysis. Two biological replicates were produced for both the fluorescent and the non-fluorescent samples.

Dataset Information

Phosphoproteomic analysis of Protein Phosphatase PfPPM2/PP2C mutant of Plasmodium falciparum 3D7

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