Top down proteomics reveals mature proteoforms expressed in subcellular fractions of the Echinococcus granulosus preadult stage

Karina R. Lorenzatto, Kyunggon Kim, Ioanna Ntai, Gabriela P. Paludo, Jeferson Camargo De Lima, Paul M. Thomas, Neil L. Kelleher, Henrique B. Ferreira*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Echinococcus granulosus is the causative agent of cystic hydatid disease, a neglected zoonosis responsible for high morbidity and mortality. Several molecular mechanisms underlying parasite biology remain poorly understood. Here, E. granulosus subcellular fractions were analyzed by top down and bottom up proteomics for protein identification and characterization of co-translational and post-translational modifications (CTMs and PTMs, respectively). Nuclear and cytosolic extracts of E. granulosus protoscoleces were fractionated by 10% GELFrEE and proteins under 30 kDa were analyzed by LC-MS/MS. By top down analysis, 186 proteins and 207 proteoforms were identified, of which 122 and 52 proteoforms were exclusively detected in nuclear and cytosolic fractions, respectively. CTMs were evident as 71% of the proteoforms had methionine excised and 47% were N-terminal acetylated. In addition, in silico internal acetylation prediction coupled with top down MS allowed the characterization of 9 proteins differentially acetylated, including histones. Bottom up analysis increased the overall number of identified proteins in nuclear and cytosolic fractions to 154 and 112, respectively. Overall, our results provided the first description of the low mass proteome of E. granulosus subcellular fractions and highlighted proteoforms with CTMs and PTMS whose characterization may lead to another level of understanding about molecular mechanisms controlling parasitic flatworm biology.

Original languageEnglish (US)
Pages (from-to)4805-4814
Number of pages10
JournalJournal of Proteome Research
Volume14
Issue number11
DOIs
StatePublished - Nov 6 2015

Keywords

  • hydatid disease
  • low mass proteome
  • post-translational modifications
  • subcellular fractions
  • top down proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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