Abstract
Sporozoites of the malaria parasite Plasmodium are transmitted by mosquitoes and infect the liver for an initial and obligatory round of replication, before exponential multiplication in the blood and onset of the disease. Sporozoites and liver stages provide attractive targets for malaria vaccines and prophylactic drugs. In this context, defining the parasite proteome is important to explore the parasite biology and to identify potential targets for antimalarial strategies. Previous studies have determined the total proteome of sporozoites from the two main human malaria parasites, P. falciparum and P. vivax, as well as P. yoelii, which infects rodents. Another murine malaria parasite, P. berghei, is widely used to investigate the parasite biology. However, a deep view of the proteome of P. berghei sporozoites is still missing. To fill this gap, we took advantage of the highly sensitive timsTOF PRO mass spectrometer, combined with three alternative methods for sporozoite purification, to identify the proteome of P. berghei sporozoites using low numbers of parasites. This study provides a reference proteome for P. berghei sporozoites, identifying a core set of proteins expressed across species, and illustrates how the unprecedented sensitivity of the timsTOF PRO system enables deep proteomic analysis from limited sample amounts.
Highlights
Malaria, with more than 200 million estimated cases and 400,000 deaths every year [1], remains a major public health problem in many countries
By employing a highly sensitive mass spectrometry approach based on trapped ion mobility spectrometry with parallel accumulation-serial fragmentation, we identified the deep proteome of P. berghei sporozoites using unprecedented low numbers of parasites, and identified a core set of sporozoite proteins expressed across Plasmodium species
Plasmodium sporozoites are obtained by hand dissection of the salivary glands of infected mosquitoes, resulting in sample contamination with proteins from the mosquito or its microbiota
Summary
With more than 200 million estimated cases and 400,000 deaths every year [1], remains a major public health problem in many countries. Motile sporozoites traffic to the liver through the blood stream, and reach the liver parenchyma where they invade hepatocytes for an initial and obligatory replication phase, resulting in the release of tens of thousands of merozoites [2] These merozoites invade erythrocytes and initiate the exponential asexual reproduction of the parasite in erythrocytes, causing the symptomatic phase of malaria. This emphasizes the need to develop more efficacious malaria vaccines targeting other antigens In this context, proteomic studies of sporozoites, by identifying the set of proteins expressed by these forms, can guide the downselection of potential new candidates. Methods have been developed to purify sporozoites and limit the proportion of mosquito proteins [5] These approaches, combined with LC-MS/MS, led to the characterization of the total proteome of P. falciparum and P. vivax sali-
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