A parasite cysteine protease is key to host protein degradation and iron acquisition.

Abstract

Cysteine proteases of the Clan CA (papain) family are the predominant protease group in primitive invertebrates. Cysteine protease inhibitors arrest infection by the protozoan parasite, Trypanosoma brucei. RNA interference studies implicated a cathepsin B-like protease, tbcatB, as a key inhibitor target. Utilizing parasites in which one of the two alleles of tbcatb has been deleted, the key role of this protease in degradation of endocytosed host proteins is delineated. TbcatB deficiency results in a decreased growth rate and dysmorphism of the flagellar pocket and the subjacent endocytic compartment. Western blot and microscopic analysis indicate that deficiency in tbcatB results in accumulation of both host and parasite proteins, including the lysosomal marker p67. A critical function for parasitism is the degradation of host transferrin, which is necessary for iron acquisition. Substrate specificity analysis of recombinant tbcatB revealed the optimal peptide cleavage sequences for the enzyme and these were confirmed experimentally using FRET-based substrates. Degradation of transferrin was validated by SDS-PAGE and the specific cleavage sites identified by N-terminal sequencing. Because even a modest deficiency in tbcatB is lethal for the parasite, tbcatB is a logical target for the development of new anti-trypanosomal chemotherapy.

• Authors: Theresa C O'Brien, Zachary B Mackey, Richard D Fetter, Youngchool Choe, Anthony J O'Donoghue, Min Zhou, Charles S Craik, Conor R Caffrey, James H McKerrow
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 283 Issue 43 Pg. 28934-43 (Oct 24 2008) ISSN: 0021-9258 [Print] United States
• PMID: 18701454 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Transferrin
• Iron
• Cysteine Endopeptidases
• TbcatB protein, Trypanosoma brucei
• Cysteine

Topics

• Animals
• Animals, Genetically Modified
• Catalytic Domain
• Cysteine (chemistry)
• Cysteine Endopeptidases (chemistry, metabolism, physiology)
• Endocytosis
• Fluorescence Resonance Energy Transfer
• Iron (chemistry, metabolism)
• Lysosomes (metabolism)
• Microscopy, Electron, Transmission
• Models, Biological
• Protein Structure, Tertiary
• RNA Interference
• Transferrin (chemistry)
• Trypanosoma brucei brucei (metabolism)