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Proteomic profiling of Plasmodium sporozoite maturation identifies new proteins essential for parasite development and infectivity.

Lasonder E, Janse CJ, van Gemert GJ, Mair GR, Vermunt AM, Douradinha BG, van Noort V, Huynen MA, Luty AJ, Kroeze H, Khan SM, Sauerwein RW, Waters AP, Mann M, Stunnenberg HG - PLoS Pathog. (2008)

Bottom Line: The different sporozoite proteomes contain a large number of stage specific proteins whose annotation suggest an involvement in sporozoite maturation, motility, infection of the human host and associated metabolic adjustments.Analyses of proteins identified in the P. falciparum sporozoite proteomes by orthologous gene disruption in the rodent malaria parasite, P. berghei, revealed three previously uncharacterized Plasmodium proteins that appear to be essential for sporozoite development at distinct points of maturation in the mosquito.This study sheds light on the development and maturation of the malaria parasite in an Anopheles mosquito and also identifies proteins that may be essential for sporozoite infectivity to humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, NCMLS, Radboud University Nijmegen, Nijmegen, The Netherlands.

ABSTRACT
Plasmodium falciparum sporozoites that develop and mature inside an Anopheles mosquito initiate a malaria infection in humans. Here we report the first proteomic comparison of different parasite stages from the mosquito -- early and late oocysts containing midgut sporozoites, and the mature, infectious salivary gland sporozoites. Despite the morphological similarity between midgut and salivary gland sporozoites, their proteomes are markedly different, in agreement with their increase in hepatocyte infectivity. The different sporozoite proteomes contain a large number of stage specific proteins whose annotation suggest an involvement in sporozoite maturation, motility, infection of the human host and associated metabolic adjustments. Analyses of proteins identified in the P. falciparum sporozoite proteomes by orthologous gene disruption in the rodent malaria parasite, P. berghei, revealed three previously uncharacterized Plasmodium proteins that appear to be essential for sporozoite development at distinct points of maturation in the mosquito. This study sheds light on the development and maturation of the malaria parasite in an Anopheles mosquito and also identifies proteins that may be essential for sporozoite infectivity to humans.

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Phenotypic characterization of P. berghei mutants (841cl1, 843cl1, 802cl1) with disrupted genes.(A) Numbers of oocyst-derived sporozoites and salivary gland sporozoites per mosquito from day 14 till day 27 post mosquito infection. Scale bars in Figure 3a indicate 50 µm. Wild type (WT) sporozoite numbers are shown in blue bars, 841 clone (PB000251.01.0/PFD0425w) gene disruptant sporozoite numbers in purple, 843 clone (PB402680.00.0/MAL8P1.66) gene disruptant sporozoite numbers in yellow, and 802 clone (PB101363.00.0-PB000829.02.0-PB105739.00.0/PF14_0435) gene disruptant sporozoite numbers are shown in pale blue. (B) Oocysts and sporozoites of the three mutant lines. Upper panel: GFP-expressing mature oocysts at day 10 after infection. Middle panel: Representative images (phase contrast microscopy) of mature (day 12) oocyst. Sporozoite formation in mutant 841 (PB000251.01.0/PFD0425w) is same as WT whereas in lines 843 (PB402680.00.0/MAL8P1.66) and 802 (PB101363.00.0-PB000829.02.0-PB105739.00.0/PF14_0435) sporozoite development is either affected (i.e. 843) or completely absent (i.e. 802). Lower panel: GFP-expressing sporozoites (released by mechanical rupture of oocysts at day 18–20). Scale bars in Figure 3b indicate 12 um. (C) Hepatocyte traversal and invasion of oocyst derived sporozoites (841, PB000251.01.0/PFD0425w)) and salivary gland sporozoites (843, (PB402680.00.0/MAL8P1.66)) compared to WT sporozoites similarly mechanically liberated from oocyst. Bars represent the average percentage of HepG2 cell traversal and invasion relative to wild type. Scale bars in Figure 3c indicate 12 um.
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ppat-1000195-g003: Phenotypic characterization of P. berghei mutants (841cl1, 843cl1, 802cl1) with disrupted genes.(A) Numbers of oocyst-derived sporozoites and salivary gland sporozoites per mosquito from day 14 till day 27 post mosquito infection. Scale bars in Figure 3a indicate 50 µm. Wild type (WT) sporozoite numbers are shown in blue bars, 841 clone (PB000251.01.0/PFD0425w) gene disruptant sporozoite numbers in purple, 843 clone (PB402680.00.0/MAL8P1.66) gene disruptant sporozoite numbers in yellow, and 802 clone (PB101363.00.0-PB000829.02.0-PB105739.00.0/PF14_0435) gene disruptant sporozoite numbers are shown in pale blue. (B) Oocysts and sporozoites of the three mutant lines. Upper panel: GFP-expressing mature oocysts at day 10 after infection. Middle panel: Representative images (phase contrast microscopy) of mature (day 12) oocyst. Sporozoite formation in mutant 841 (PB000251.01.0/PFD0425w) is same as WT whereas in lines 843 (PB402680.00.0/MAL8P1.66) and 802 (PB101363.00.0-PB000829.02.0-PB105739.00.0/PF14_0435) sporozoite development is either affected (i.e. 843) or completely absent (i.e. 802). Lower panel: GFP-expressing sporozoites (released by mechanical rupture of oocysts at day 18–20). Scale bars in Figure 3b indicate 12 um. (C) Hepatocyte traversal and invasion of oocyst derived sporozoites (841, PB000251.01.0/PFD0425w)) and salivary gland sporozoites (843, (PB402680.00.0/MAL8P1.66)) compared to WT sporozoites similarly mechanically liberated from oocyst. Bars represent the average percentage of HepG2 cell traversal and invasion relative to wild type. Scale bars in Figure 3c indicate 12 um.

Mentions: See Figure 3 for details of the characterization of the 3 mutants with a phenotype different from wild type.


Proteomic profiling of Plasmodium sporozoite maturation identifies new proteins essential for parasite development and infectivity.

Lasonder E, Janse CJ, van Gemert GJ, Mair GR, Vermunt AM, Douradinha BG, van Noort V, Huynen MA, Luty AJ, Kroeze H, Khan SM, Sauerwein RW, Waters AP, Mann M, Stunnenberg HG - PLoS Pathog. (2008)

Phenotypic characterization of P. berghei mutants (841cl1, 843cl1, 802cl1) with disrupted genes.(A) Numbers of oocyst-derived sporozoites and salivary gland sporozoites per mosquito from day 14 till day 27 post mosquito infection. Scale bars in Figure 3a indicate 50 µm. Wild type (WT) sporozoite numbers are shown in blue bars, 841 clone (PB000251.01.0/PFD0425w) gene disruptant sporozoite numbers in purple, 843 clone (PB402680.00.0/MAL8P1.66) gene disruptant sporozoite numbers in yellow, and 802 clone (PB101363.00.0-PB000829.02.0-PB105739.00.0/PF14_0435) gene disruptant sporozoite numbers are shown in pale blue. (B) Oocysts and sporozoites of the three mutant lines. Upper panel: GFP-expressing mature oocysts at day 10 after infection. Middle panel: Representative images (phase contrast microscopy) of mature (day 12) oocyst. Sporozoite formation in mutant 841 (PB000251.01.0/PFD0425w) is same as WT whereas in lines 843 (PB402680.00.0/MAL8P1.66) and 802 (PB101363.00.0-PB000829.02.0-PB105739.00.0/PF14_0435) sporozoite development is either affected (i.e. 843) or completely absent (i.e. 802). Lower panel: GFP-expressing sporozoites (released by mechanical rupture of oocysts at day 18–20). Scale bars in Figure 3b indicate 12 um. (C) Hepatocyte traversal and invasion of oocyst derived sporozoites (841, PB000251.01.0/PFD0425w)) and salivary gland sporozoites (843, (PB402680.00.0/MAL8P1.66)) compared to WT sporozoites similarly mechanically liberated from oocyst. Bars represent the average percentage of HepG2 cell traversal and invasion relative to wild type. Scale bars in Figure 3c indicate 12 um.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2570797&req=5

ppat-1000195-g003: Phenotypic characterization of P. berghei mutants (841cl1, 843cl1, 802cl1) with disrupted genes.(A) Numbers of oocyst-derived sporozoites and salivary gland sporozoites per mosquito from day 14 till day 27 post mosquito infection. Scale bars in Figure 3a indicate 50 µm. Wild type (WT) sporozoite numbers are shown in blue bars, 841 clone (PB000251.01.0/PFD0425w) gene disruptant sporozoite numbers in purple, 843 clone (PB402680.00.0/MAL8P1.66) gene disruptant sporozoite numbers in yellow, and 802 clone (PB101363.00.0-PB000829.02.0-PB105739.00.0/PF14_0435) gene disruptant sporozoite numbers are shown in pale blue. (B) Oocysts and sporozoites of the three mutant lines. Upper panel: GFP-expressing mature oocysts at day 10 after infection. Middle panel: Representative images (phase contrast microscopy) of mature (day 12) oocyst. Sporozoite formation in mutant 841 (PB000251.01.0/PFD0425w) is same as WT whereas in lines 843 (PB402680.00.0/MAL8P1.66) and 802 (PB101363.00.0-PB000829.02.0-PB105739.00.0/PF14_0435) sporozoite development is either affected (i.e. 843) or completely absent (i.e. 802). Lower panel: GFP-expressing sporozoites (released by mechanical rupture of oocysts at day 18–20). Scale bars in Figure 3b indicate 12 um. (C) Hepatocyte traversal and invasion of oocyst derived sporozoites (841, PB000251.01.0/PFD0425w)) and salivary gland sporozoites (843, (PB402680.00.0/MAL8P1.66)) compared to WT sporozoites similarly mechanically liberated from oocyst. Bars represent the average percentage of HepG2 cell traversal and invasion relative to wild type. Scale bars in Figure 3c indicate 12 um.
Mentions: See Figure 3 for details of the characterization of the 3 mutants with a phenotype different from wild type.

Bottom Line: The different sporozoite proteomes contain a large number of stage specific proteins whose annotation suggest an involvement in sporozoite maturation, motility, infection of the human host and associated metabolic adjustments.Analyses of proteins identified in the P. falciparum sporozoite proteomes by orthologous gene disruption in the rodent malaria parasite, P. berghei, revealed three previously uncharacterized Plasmodium proteins that appear to be essential for sporozoite development at distinct points of maturation in the mosquito.This study sheds light on the development and maturation of the malaria parasite in an Anopheles mosquito and also identifies proteins that may be essential for sporozoite infectivity to humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, NCMLS, Radboud University Nijmegen, Nijmegen, The Netherlands.

ABSTRACT
Plasmodium falciparum sporozoites that develop and mature inside an Anopheles mosquito initiate a malaria infection in humans. Here we report the first proteomic comparison of different parasite stages from the mosquito -- early and late oocysts containing midgut sporozoites, and the mature, infectious salivary gland sporozoites. Despite the morphological similarity between midgut and salivary gland sporozoites, their proteomes are markedly different, in agreement with their increase in hepatocyte infectivity. The different sporozoite proteomes contain a large number of stage specific proteins whose annotation suggest an involvement in sporozoite maturation, motility, infection of the human host and associated metabolic adjustments. Analyses of proteins identified in the P. falciparum sporozoite proteomes by orthologous gene disruption in the rodent malaria parasite, P. berghei, revealed three previously uncharacterized Plasmodium proteins that appear to be essential for sporozoite development at distinct points of maturation in the mosquito. This study sheds light on the development and maturation of the malaria parasite in an Anopheles mosquito and also identifies proteins that may be essential for sporozoite infectivity to humans.

Show MeSH
Related in: MedlinePlus