Interrogating the Plasmodium Sporozoite Surface

Identification of Surface-Exposed Proteins and Demonstration of Glycosylation on CSP and TRAP by Mass Spectrometry-Based Proteomics

Kristian E. Swearingen, Scott E. Lindner, Lirong Shi, Melanie J. Shears, Anke Harupa, Christine S. Hopp, Ashley M. Vaughan, Timothy A. Springer, Robert L. Moritz, Stefan H I Kappe, Photini Sinnis

Research output: Contribution to journalArticle

Abstract

Malaria parasite infection is initiated by the mosquito-transmitted sporozoite stage, a highly motile invasive cell that targets hepatocytes in the liver for infection. A promising approach to developing a malaria vaccine is the use of proteins located on the sporozoite surface as antigens to elicit humoral immune responses that prevent the establishment of infection. Very little of the P. falciparum genome has been considered as potential vaccine targets, and candidate vaccines have been almost exclusively based on single antigens, generating the need for novel target identification. The most advanced malaria vaccine to date, RTS,S, a subunit vaccine consisting of a portion of the major surface protein circumsporozoite protein (CSP), conferred limited protection in Phase III trials, falling short of community-established vaccine efficacy goals. In striking contrast to the limited protection seen in current vaccine trials, sterilizing immunity can be achieved by immunization with radiation-attenuated sporozoites, suggesting that more potent protection may be achievable with a multivalent protein vaccine. Here, we provide the most comprehensive analysis to date of proteins located on the surface of or secreted by Plasmodium falciparum salivary gland sporozoites. We used chemical labeling to isolate surface-exposed proteins on sporozoites and identified these proteins by mass spectrometry. We validated several of these targets and also provide evidence that components of the inner membrane complex are in fact surface-exposed and accessible to antibodies in live sporozoites. Finally, our mass spectrometry data provide the first direct evidence that the Plasmodium surface proteins CSP and TRAP are glycosylated in sporozoites, a finding that could impact the selection of vaccine antigens.

Original languageEnglish (US)
Article numbere1005606
JournalPLoS Pathogens
Volume12
Issue number4
DOIs
StatePublished - Apr 1 2016

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Sporozoites
Plasmodium
Glycosylation
Proteomics
Mass Spectrometry
Membrane Proteins
Vaccines
Proteins
Malaria Vaccines
Antigens
Parasitic Diseases
Subunit Vaccines
Plasmodium falciparum
Surface Antigens
Humoral Immunity
Infection
Salivary Glands
Culicidae
Malaria
Hepatocytes

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

Interrogating the Plasmodium Sporozoite Surface : Identification of Surface-Exposed Proteins and Demonstration of Glycosylation on CSP and TRAP by Mass Spectrometry-Based Proteomics. / Swearingen, Kristian E.; Lindner, Scott E.; Shi, Lirong; Shears, Melanie J.; Harupa, Anke; Hopp, Christine S.; Vaughan, Ashley M.; Springer, Timothy A.; Moritz, Robert L.; Kappe, Stefan H I; Sinnis, Photini.

In: PLoS Pathogens, Vol. 12, No. 4, e1005606, 01.04.2016.

Research output: Contribution to journalArticle

Swearingen, Kristian E. ; Lindner, Scott E. ; Shi, Lirong ; Shears, Melanie J. ; Harupa, Anke ; Hopp, Christine S. ; Vaughan, Ashley M. ; Springer, Timothy A. ; Moritz, Robert L. ; Kappe, Stefan H I ; Sinnis, Photini. / Interrogating the Plasmodium Sporozoite Surface : Identification of Surface-Exposed Proteins and Demonstration of Glycosylation on CSP and TRAP by Mass Spectrometry-Based Proteomics. In: PLoS Pathogens. 2016 ; Vol. 12, No. 4.
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