pH regulation in glycosomes of procyclic form Trypanosoma brucei

TitlepH regulation in glycosomes of procyclic form Trypanosoma brucei
Publication TypeJournal Article
Year of Publication2017
AuthorsLin, S., Voyton C., Morris M. T., Ackroyd P. C., Morris J. C., and Christensen K. A.
JournalJ Biol Chem
Volume292
Pagination7795-7805
Date PublishedMay 12
ISBN Number1083-351X (Electronic)<br/>0021-9258 (Linking)
Accession Number28348078
Keywordsbiosensor, fluorescence, glucose metabolism, microscopy, parasite metabolism, parasitology, proton pump, Trypanosoma brucei, trypanosome
Abstract

Here we report the use of a fluorescein-tagged peroxisomal targeting sequence peptide (F-PTS1, acetyl-C{K(FITC)}GGAKL) for investigating pH regulation of glycosomes in live procyclic form Trypanosoma brucei When added to cells, this fluorescent peptide is internalized within vesicular structures, including glycosomes, and can be visualized after 30-60 min. Using F-PTS1 we are able to observe the pH conditions inside glycosomes in response to starvation conditions. Previous studies have shown that in the absence of glucose, the glycosome exhibits mild acidification from pH 7.4 +/- 0.2 to 6.8 +/- 0.2. Our results suggest that this response occurs under proline starvation as well. This pH regulation is found to be independent from cytosolic pH and requires a source of Na+ ions. Glycosomes were also observed to be more resistant to external pH changes than the cytosol; placement of cells in acidic buffers (pH 5) reduced the pH of the cytosol by 0.8 +/- 0.1 pH units, whereas glycosomal pH decreases by 0.5 +/- 0.1 pH units. This observation suggests that regulation of glycosomal pH is different and independent from cytosolic pH regulation. Furthermore, pH regulation is likely to work by an active process, because cells depleted of ATP with 2-deoxyglucose and sodium azide were unable to properly regulate pH. Finally, inhibitor studies with bafilomycin and EIPA suggest that both V-ATPases and Na+/H+ exchangers are required for glycosomal pH regulation.

URLhttps://www.jbc.org/content/292/19/7795.long
DOI10.1074/jbc.M117.784173
Alternate JournalJournal of computational chemistry