Figure 5.

Fractions of Leishmania major-derived extracellular vesicles display distinct uniquely expressed proteins. (A) The protein content of Leishmania major-derived EVs was catalogued by mass spectrometry (L. major database) using biological triplicates obtained for each fraction (n=3) and analyzed using Mascot followed by Scaffold Software. Total proteins detected by LC-MS/MS in each fraction were determined using inclusion criteria of a minimum of 2 detected peptides, a peptide threshold of 80%, and a protein identity of 95%. Uniqueness was then determined based on a true hit being identified in only one of the fractions. (B) A protein-protein interaction networks of uniquely expressed proteins in the 100K fraction was generated using unique proteins detected by LC-MS/MS in the 100K fraction of Leishmania major-derived EVs. Notable proteins include SNF-7, Qc-SNARE, IST1, SNF-7 superfamily, VTA1, CHMP2B, ALIX, and VPS37 were identified in this group, which are part of the ESCRT pathway, and PP5 and PRL-1 which are parasite phosphatases were also identified. The protein-protein interaction network was created using String database (https://www.string-db.org) using the highest confidence of interaction (>0.900), showing only connected proteins in the network and using an MCL inflation parameter of 3.