Apical Surface Expression of Aspartic Protease Plasmepsin 4, a Potential Transmission-blocking Target of the Plasmodium Ookinete*

To invade its definitive host, the mosquito, the malaria parasite must cross the midgut peritrophic matrix that is composed of chitin cross-linked by chitin-binding proteins and then develop into an oocyst on the midgut basal lamina. Previous evidence indicates that Plasmodium ookinete-secreted chitinase is important in midgut invasion. The mechanistic role of other ookinete-secreted enzymes in midgut invasion has not been previously examined. De novo mass spectrometry sequencing of a protein obtained by benzamidine affinity column of Plasmodium gallinaceum ookinete axenic culture supernatant demonstrated the presence of an ookinete-secreted plasmepsin, an aspartic protease previously only known to be present in the digestive vacuole of asexual stage malaria parasites. This plasmepsin, the ortholog of Plasmodium falciparum plasmepsin 4, was designated PgPM4. PgPM4 and PgCHT2 (the P. gallinaceum ortholog of P. falciparum chitinase PfCHT1) are both localized on the ookinete apical surface, and both are present in micronemes. Aspartic protease inhibitors (peptidomimetic and natural product), calpain inhibitors, and anti-PgPM4 monoclonal antibodies significantly reduced parasite infectivity for mosquitoes. These results suggest that plasmepsin 4, previously known only to function in the digestive vacuole of asexual blood stage Plasmodium, plays a role in how the ookinete interacts with the mosquito midgut interactions as it becomes an oocyst. These data are the first to delineate a role for an aspartic protease in mediating Plasmodium invasion of the mosquito and demonstrate the potential for plasmepsin 4 as a malaria transmission-blocking vaccine target.

• Publication year

  2010

• Venue

  Journal of Biological Chemistry

• Publication date

  2010-01-07

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1074/jbc.M109.063388PMID 20056606PMCID PMC2832958
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• Aspartic protease inhibitors, calpain inhibitors, and anti-PgPM4 monoclonal antibodies reduce parasite infectivity for mosquitoes, implicating PgPM4 in mosquito midgut invasion and transmission blocking.

  Confidence 0.94

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• PgPM4 and PgCHT2 localize to the ookinete apical surface and are present in micronemes.

  Confidence 0.96

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• De novo mass spectrometry sequencing identified an ookinete-secreted plasmepsin from Plasmodium gallinaceum axenic culture supernatant, and the protein was designated PgPM4 as the ortholog of Plasmodium falciparum plasmepsin 4.

  Confidence 0.98

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review

• apical surface

  cellular location, subcellular location

  The tip region of the ookinete membrane where invasion-associated proteins are localized in this paper.

  Aliases: ookinete apical surface

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• benzamidine affinity column

  purification method, chromatography medium

  An affinity purification matrix used to recover the protease-containing protein fraction from ookinete culture supernatant.

  Aliases: benzamidine column

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• de novo mass spectrometry sequencing

  method

  A protein identification approach used here to determine the sequence of the recovered ookinete-secreted protease.

  Aliases: de novo MS sequencing, mass spectrometry sequencing

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• micronemes

  organelle, cellular compartment

  Secretory organelles in the ookinete that contain invasion-related proteins examined in this study.

  Aliases: microneme

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• mosquito infectivity

  phenotype, outcome

  The ability of the parasite to establish infection in mosquitoes after exposure to treated ookinetes.

  Aliases: infectivity for mosquitoes

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• mosquito midgut invasion

  biological process

  The process by which the ookinete crosses the mosquito midgut barrier during infection.

  Aliases: midgut invasion

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• pgcht2

  protein, enzyme

  The Plasmodium gallinaceum ortholog of Plasmodium falciparum chitinase PfCHT1 studied alongside PgPM4.

  Aliases: Plasmodium gallinaceum chitinase 2

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• pgpm4

  protein, protease

  The Plasmodium gallinaceum plasmepsin 4 ortholog characterized from ookinete secretions in this work.

  Aliases: Plasmodium gallinaceum plasmepsin 4

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• plasmodium gallinaceum ookinete

  parasite stage, cell type

  The motile sexual-stage parasite form from Plasmodium gallinaceum that interacts with the mosquito midgut.

  Aliases: ookinete

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review
• plasmodium gallinaceum ookinete axenic culture supernatant

  sample, biological specimen

  The cell-free medium collected from cultured Plasmodium gallinaceum ookinetes that supplied the protein used for identification.

  Aliases: ookinete culture supernatant

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) review

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