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Scientists at TSRI identify thousands of proteins associated with deadliest form of malaria

like Plasmodium falciparum, "proteomics" adds the topographical information to that map by identifying which genes are actually expressed as proteins in the Plasmodium falciparum cells.

More importantly, Florens and Yates also sought to identify which proteins are expressed at which stages of the organism's lifecycle. This was no small task. Plasmodium falciparum has at least ten distinct stages in its lifecycle, and there is no way of telling which are expressed at each distinct stage of the pathogen's lifecycle simply by looking at the genes.

But Florens and Yates were able to figure out which proteins were expressed during four different stages (sporozoites, merozoites, trophozoites, and gametocytes) and, thus, which might make good vaccine targets.

Mass Spectrometry and Malaria

The process was basically to take samples of a single isolate of Plasmodium falciparum and grow three of the four different stages in blood in a way that allowed samples to be purified. The fourth stage, the sporozoites, had to be hand-dissected from mosquito salivary glands.

In purifying the samples, Yates and Florens first separated the soluble proteins from the membrane-bound proteins, then digested them (chopped into smaller "peptide" pieces with enzymes), and resolved them using liquid chromatography combined with tandem mass spectrometry.

The instrument detects the pieces and uses sophisticated software that Yates and his colleagues developed previously to search a database of predicted genes to reconstruct most of the proteins in the sample. This technique was particularly useful in this context because it allowed a very large background "noise" of mosquito and human proteins to be subtracted out. The peptides that come from the Plasmodium can be distinguished from those that come from the mosquito or the human.

Furthermore, using the technique, Florens and Yates were able to show not only which genes were expressed in eac
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Contact: Keith McKeown
kmckeown@scripps.edu
858-784-8134
Scripps Research Institute
3-Oct-2002


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