Identification of the glycosylphosphatidylinositol-specific phospholipase A2 (GPI-PLA2) that mediates GPI fatty acid remodeling in Trypanosoma brucei

Ji, Z., Nagar, R., Duncan, S. M., Sampaio Guther, M. L. and Ferguson, M. A.J. (2023) Identification of the glycosylphosphatidylinositol-specific phospholipase A2 (GPI-PLA2) that mediates GPI fatty acid remodeling in Trypanosoma brucei. Journal of Biological Chemistry, 299(8), 105016. (doi: 10.1016/j.jbc.2023.105016) (PMID:37414151) (PMCID:PMC10457582)

Abstract

The biosynthesis of glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) in the parasitic protozoan Trypanosoma brucei involves fatty acid remodeling of the GPI precursor molecules before they are transferred to protein in the endoplasmic reticulum. The genes encoding the requisite phospholipase A2 and A1 activities for this remodeling have thus far been elusive. Here, we identify a gene, Tb927.7.6110, that encodes a protein that is both necessary and sufficient for GPI-phospholipase A2 (GPI-PLA2) activity in the procyclic form of the parasite. The predicted protein product belongs to the alkaline ceramidase, PAQR receptor, Per1, SID-1, and TMEM8 (CREST) superfamily of transmembrane hydrolase proteins and shows sequence similarity to Post-GPI-Attachment to Protein 6 (PGAP6), a GPI-PLA2 that acts after transfer of GPI precursors to protein in mammalian cells. We show the trypanosome Tb927.7.6110 GPI-PLA2 gene resides in a locus with two closely related genes Tb927.7.6150 and Tb927.7.6170, one of which (Tb927.7.6150) most likely encodes a catalytically inactive protein. The absence of GPI-PLA2 in the null mutant procyclic cells not only affected fatty acid remodeling but also reduced GPI anchor sidechain size on mature GPI-anchored procyclin glycoproteins. This reduction in GPI anchor sidechain size was reversed upon the re-addition of Tb927.7.6110 and of Tb927.7.6170, despite the latter not encoding GPI precursor GPI-PLA2 activity. Taken together, we conclude that Tb927.7.6110 encodes the GPI-PLA2 of GPI precursor fatty acid remodeling and that more work is required to assess the roles and essentiality of Tb927.7.6170 and the presumably enzymatically inactive Tb927.7.6150.

• Item Type: Articles
• Additional Information: This work was supported by a China Scholarship Council PhD scholarship to Z. J. (201706310166) and a Wellcome Investigator Award to M. A. J. F. (101842/Z13/Z).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Duncan, Mr Samuel

Creator Roles

Duncan, S.: Writing – original draft, Supervision

• Authors: Ji, Z., Nagar, R., Duncan, S. M., Sampaio Guther, M. L., and Ferguson, M. A.J.
• College/School: College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
• Journal Name: Journal of Biological Chemistry
• Publisher: Elsevier
• ISSN: 0021-9258
• ISSN (Online): 1083-351X
• Published Online: 05 July 2023
• Copyright Holders: Copyright © 2023 The Authors
• First Published: First published in Journal of Biological Chemistry 299(8):105016
• Publisher Policy: Reproduced under a Creative Commons license