Is dihydrolipoic acid among the reductive activators of parasite CysHis proteases?
Activities of mature CysHis proteases depend upon relative rates of oxidations vs. reductions of catalytic sulfur by multiple enzymatic and non-enzymatic reactions. CysHis peptidolysis is inhibited by Fe3+ but not Fe2+. Others report the paradox that malarial parasites require exogenous free lipoic acid (LA) from human host, although the apicoplast organelle produces it. Extra-cellular LA disulfide can be taken up and reduced to dihydrolipoic acid (DHLA) by reductases of any cell type. Here, the opposing effects of DHLA vs. Fe3+ on the falcipain-2 hemoglobinase were investigated employing Z-Phe-Arg-AMC substrate. Despite limited solubility, non-regenerated DHLA (10 microM, threshold 2 microM) was found to be the most potent activator of the air-inactivated (sulfoxygenated) protease discovered thus far. Activation was preemptively opposed by Fe3+, but not Fe2+. However, cruzain from T. cruzi, and cathepsin B from mammal were indistinguishable in their responsiveness to DHLA and Fe redox. Thus, DHLA activation vs. Fe3+ inhibition is not unique to falcipain-2 or apicomplexans but is rather a primordial feature of CysHis peptidolysis. Free LA and/or unassociated lipoylated enzyme subunits could be among multiple pathways shuttling reducing equivalents to reduction of proteins, including CysHis proteases. It is discussed that opposing DHLA-Fe3+ modification of plasmodial proteolysis might be a specialized adaptation to intra-erythrocytic growth.
Lockwood
Thomas D
TD
Department of Pharmacology and Toxicology, School of Medicine, Wright State University, Dayton, OH 45435, USA. thomas.lockwood@wright.edu
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002401:Cathepsin B / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004793:Enzyme Reactivators / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D006639:Histidine / Q000378:metabolism; D007501:Iron / Q000494:pharmacology; D008322:Mammals; D010084:Oxidation-Reduction; D010963:Plasmodium falciparum / Q000201:enzymology; D015800:Protozoan Proteins / Q000378:metabolism; D008063:Thioctic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000201:enzymology
D000975:Antioxidants; D004793:Enzyme Reactivators; D015800:Protozoan Proteins; D006639:Histidine; D008063:Thioctic Acid; C015144:dihydrolipoic acid; D007501:Iron; D003546:Cysteine Endopeptidases; C074032:cruzain, Trypanosoma cruzi; C416224:falcipain 2; D002401:Cathepsin B; D005978:Glutathione; D003545:Cysteine; D004229:Dithiothreitol