ATP production in isolated mitochondria of procyclic Trypanosoma brucei.

This chapter describes a luciferase-based protocol to measure adenosine triphosphate (ATP) production in isolated mitochondria of Trypanosoma brucei. The assay represents an excellent method to characterize the functionality of isolated mitochondria. Comparing the ATP production induced by substrates for oxidative phosphorylation to the one induced by substrates for substrate-level phosphorylation allows conclusions regarding the integrity of the outer and inner mitochondrial membranes.

Elongation factor 1a mediates the specificity of mitochondrial tRNA import in T. brucei.

Mitochondrial tRNA import is widespread in eukaryotes. Yet, the mechanism that determines its specificity is unknown. Previous in vivo experiments using the tRNAs(Met), tRNA(Ile) and tRNA(Lys) have suggested that the T-stem nucleotide pair 51:63 is the main localization determinant of tRNAs in Trypanosoma brucei.

Type 3 peptide deformylases are required for oxidative phosphorylation in Trypanosoma brucei.

Peptide deformylase (PDF) catalyses the removal of the formyl group from the first methionine of nascent proteins. Type 1 PDFs are found in bacteria and have orthologues in most eukaryotes. Type 2 PDFs are restricted to bacteria.