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Mitochondrial alternative pathways: biochemical and molecular studies of an UCP-like from Aspergillus sp


We previously characterized the mitochondrial respiratory chain of A. fumigatus. We have shown that, in addition to the cytochrome c pathway, this organism possesses alternative pathways, involving an alternative oxidase, NADH dehydrogenase and uncoupling protein. These pathways have been associated with both energetic and antioxidant defense of cells, and represents an important factor for the fungal survival in macrophages during host invasion. There are many evidences of the existence of UCP in microorganism such as Candida parasiplosis, Acanthamoeba castelani and Aspergillus sp. However, so far any DNA sequence for UCP was described in these microorganisms. Preliminary results in our lab using degenerated primers design from multiple alignments of the deduced amino-acid sequences of UCP proteins, we got a sequence, wich contains three of molecular signatures for UCPs. The aim this project is cloning, sequencing and heterologous expression in E. coli of an UCP-like sequence. In addition, the recombinant protein will be reconstituted in proteoliposomes for biochemical characterization. (AU)

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