Dataset Information

FLAG-tag based co-immunoprecipitation of the Plasmodium falciparum Pfs230:Pfs48/45 complex

ABSTRACT: Malaria is a devastating disease, causing over half a million deaths annually mostly amongst young children. Transmission blocking vaccines aim to induce immunity against the sexual stages of the causative Plasmodium parasites, preventing human-to-mosquito transmission. Two promising transmission blocking vaccine antigens, Pfs230 and Pfs48/45, form a complex together on the surface of sexual-stage malaria parasites. We created a transgenic parasite line (iGP2230-tag) where the pfs230 gene is fused to a coding sequence for a C-terminal affinity-tag (linker-3xFLAG-linker-EPEA*). Using FLAG-tag resin, we co-immunoprecipitated the Pfs230-FLAG protein from mature gametocytes to identify components of the Pfs230:Pfs48/45 complex.

INSTRUMENT(S):

ORGANISM(S): Plasmodium Falciparum

TISSUE(S): Cell Culture

SUBMITTER: Dick Zijlmans

LAB HEAD: Matthijs Jore

PROVIDER: PXD060716 | Pride | 2025-10-09

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20240305_Exploris1_EXT_EB_EBF2d_gmctV_antiFLAG_2D_1.raw	Raw
	20240305_Exploris1_EXT_EB_EBF2d_gmctV_antiFLAG_2D_2.raw	Raw
	20240305_Exploris1_EXT_EB_EBF2d_gmctV_antiFLAG_2D_3.raw	Raw
	20240305_Exploris1_EXT_EB_EBF2d_gmctV_antiFLAG_2D_txt.zip	Other
	20240305_Exploris1_EXT_EB_NF54_WT_gmctV_antiFLAG_WT_1.raw	Raw

Items per page:

1 - 5 of 8

Similar Datasets

Project description:For malaria transmission, the parasite must undergo sexual differentiation into mature gametocytes. However, the molecular basis for this critical transition in the parasites life cycle is unknown. Six previously uncharacterized genes, Pfg14.744, Pfg14.745, Pfg14.748, Pfg14.763, Pfg14.752 and Pfg6.6 that are members of a 36 gene Plasmodium falciparum-specific subtelomeric superfamily were found to be expressed in parasites that are committed to sexual development as suggested by co-expression of Pfs16 and Pfg27. Northern blots demonstrated that Pfg14.744 and Pfg14.748 were first expressed before the parasites differentiated into morphologically distinct gametocytes, transcription continued to increase until stage II gametocytes were formed and then rapidly decreased. Immunofluorescence assays indicated that both proteins were only produced in the subpopulation of ring stage parasites that are committed to gametocytogenesis and both localized to the parasitophorous vacuole (PV)b of the early ring stage parasites. As the parasites continued to develop Pfg14.748 remained within the parasitophorous vacuole, while Pfg14.744 was detected in the erythrocyte. The 5' flanking region of either gene alone was sufficient to drive early gametocyte specific expression of green fluorescent protein (GFP). In parasites transfected with a plasmid containing the Pfg14.748 5' flanking region immediately upstream of GFP, fluorescence was observed in a small number of schizonts the cycle before stage I gametocytes were observed. This expression pattern is consistent with commitment to sexual differentiation prior to merozoite release and erythrocyte invasion. Further investigation into the role of these genes in the transition from asexual to sexual differentiation could provide new strategies to block malaria transmission. Microarray analysis was used to compare two clones derived from Plasmodium falciparum strain 3D7 parasites that differ in their ability to undergo gametocytogenesis. Clone G+ produces gametocytes and clone G- produces very few if any gametocytes. RNA was harvested from the cultures when the asexual parasitemia was 0.9-1.48% (day 4) (n=4) after setting up the gametocyte cultures and 5.2-5.58% (day 6) (n=4) prior to the appearance of morphologically distinct gametocytes and used to generate cDNA that was labeled with Cy3 or Cy5 and hybridized to the Plasmodium falciparum 70 mer oligonucleotide microarray developed by DeRisi and co-workers.

Dataset Information

FLAG-tag based co-immunoprecipitation of the Plasmodium falciparum Pfs230:Pfs48/45 complex

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets