The hydrophobic region of the Leishmania peroxin 14 : requirements for association with a glycosome mimetic membrane
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Protein import into the Leishmania glycosome requires docking of the cargo-loaded peroxin 5 (PEX5) receptor to the peroxin 14 (PEX14) bound to the glycosome surface. To examine the LdPEX14-membrane interaction, we purified L. donovani promastigote glycosomes and determined the phospholipid and fatty acid composition. These membranes contained predominately phosphatidylethanolamine, phosphatidylcholine, and phosphatidylglycerol (PG) modified primarily with C18 and C22 unsaturated fatty acid. Using large unilamellar vesicles (LUVs) with a lipid composition mimicking the glycosomal membrane in combination with sucrose density centrifugation and fluorescence-activated cell sorting technique, we established that the LdPEX14 membrane-binding activity was dependent on a predicted transmembrane helix found within residues 149-179. Monolayer experiments showed that the incorporation of PG and phospholipids with unsaturated fatty acids, which increase membrane fluidity and favor a liquid expanded phase, facilitated the penetration of LdPEX14 into biological membranes. Moreover, we demonstrated that the binding of LdPEX5 receptor or LdPEX5-PTS1 receptor-cargo complex was contingent on the presence of LdPEX14 at the surface of LUVs.
Cyr , N , Smith , T K , Boisselier , É , Leroux , L-P , Kottarampatel , A H , Davidsen , A , Salesse , C & Jardim , A 2018 , ' The hydrophobic region of the Leishmania peroxin 14 : requirements for association with a glycosome mimetic membrane ' , Biochemical Journal , vol. 475 , no. 2 , pp. 511-529 . https://doi.org/10.1042/BCJ20170746
© 2018, the Author(s). This work has been made available online in accordance with the publisher’s policies. This is the author created, accepted version manuscript following peer review and may differ slightly from the final published version. The final published version of this work is available at https://doi.org/10.1042/BCJ20170746
DescriptionThis work was funded by operating grants from the Canadian Institutes of Health Research (CIHR) and a Natural Sciences Engineering Research Council of Canada (NSERC) Discovery grant [Fonds de recherche du Québec — Nature et technologies (FRQNT) Regroupement Stratégique grant to the Centre for Host-Parasite Interactions (A.J.)]. N.C. was supported by a doctoral research scholarship from FRQNT. E.B. was supported by a Banting postdoctoral fellowship from CIHR. This work was also supported in part by Wellcome Trust grants [086658 and 093228] to T.K.S. C.S. recognizes the financial support from the Natural Sciences and Engineering Research Council of Canada and a Canada Foundation for Innovation grant .
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