Reference and Ghost Proteins Identification in Rat C6 Glioma Extracellular Vesicles

Adriana-Natalia Murgoci; Tristan Cardon; Soulaimane Aboulouard; Marie Duhamel; Isabelle Fournier; Dasa Cizkova; Michel Salzet

doi:10.1016/j.isci.2020.101045

Reference and Ghost Proteins Identification in Rat C6 Glioma Extracellular Vesicles

iScience. 2020 May 22;23(5):101045. doi: 10.1016/j.isci.2020.101045. Epub 2020 Apr 10.

Authors

Adriana-Natalia Murgoci¹, Tristan Cardon², Soulaimane Aboulouard², Marie Duhamel², Isabelle Fournier², Dasa Cizkova³, Michel Salzet⁴

Affiliations

¹ Univ. Lille, Inserm, U-1192 - Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse-PRISM, Lille 59000, France; Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava 84510, Slovakia.
² Univ. Lille, Inserm, U-1192 - Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse-PRISM, Lille 59000, France.
³ Univ. Lille, Inserm, U-1192 - Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse-PRISM, Lille 59000, France; Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava 84510, Slovakia; Department of Anatomy, Histology and Physiology, University of Veterinary Medicine and Pharmacy in Košice, Košice 04181, Slovakia. Electronic address: dasa.cizkova@uvlf.sk.
⁴ Univ. Lille, Inserm, U-1192 - Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse-PRISM, Lille 59000, France. Electronic address: michel.salzet@univ-lille.fr.

Abstract

Extracellular vesicles (EVs) mediate intercellular communication and regulate a broad range of biological processes. Novel therapeutic strategies have emerged based on the use of EVs as biological nanoparticles. To separate isolated EVs from protein aggregates and the external part of EVs membrane proteins, we performed a Trypsin/Lys C digestion treatment of EVs pellets, followed by Amicon filtration. After these steps, all the fractions have been subjected to proteomic analyses. Comparison between 6 h Trypsin/Lys C treatment or non-treated EVs revealed a quantitative variation of the surface proteins. Some surface proteins have been demasked after 6 h enzymatic digestion like CD81, CD82, Ust, Vcan, Lamp 1, Rab43, Annexin A2, Synthenin, and VSP37b. Moreover, six ghost proteins have also been identified and one corresponds to a long noncoding RNA. We thus demonstrate the presence of ghost proteins in EVs produced by glioma cells that can contribute to tumorigenesis.

Keywords: Biochemistry; Cancer; Proteomics.