The Leishmania mexicana proteasome.
As a start to understanding the importance of intracellular proteolysis in the protozoon Leishmania mexicana, the parasite proteasome has been purified and characterised. The L. mexicana proteasome is similar to proteasomes from other eukaryotes. It is soluble, and the 20S form has a mass of around 670 kDa, composed of at least 10 distinct subunits in the 22 to 32 kDa size range. The molecular mass of the L. mexicana proteasome increases to 1200 kDa in the presence of adenosine-5'-triphosphate, consistent with there being a 26S proteasome in the parasite. The purified 20S proteasome has activity towards substrates with hydrophobic, basic and acidic P, residues, and is sensitive to a range of peptide aldehyde inhibitors, as well as the proteasome-specific inhibitor lactacystin. The peptide aldehydes are able to arrest parasite growth in vitro with the same relative effectiveness as against the purified proteasome activity. The parasite population arrests with an increased 4N DNA content, indicating that, in part, the essential nature of the proteasome for L. mexicana proliferation is due to a role in the parasite cell cycle. Surprisingly, lactacystin is a relatively inefficient inhibitor of L. mexicana growth in vitro.
Robertson
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Wellcome Unit of Molecular Parasitology, The Anderson College, University of Glasgow, UK.
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003374:Coumarins; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D007894:Leishmania mexicana / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D009842:Oligopeptides; D046988:Proteasome Endopeptidase Complex; D013379:Substrate Specificity
D003374:Coumarins; D015853:Cysteine Proteinase Inhibitors; D009097:Multienzyme Complexes; D009842:Oligopeptides; C067713:lactacystin; D000255:Adenosine Triphosphate; C046582:succinyl-leucyl-leucyl-valyl-tyrosyl-methylcoumarinamide; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D000111:Acetylcysteine