High-Efficiency Isolation of Nuclear Envelope Protein Complexes from Trypanosomes

Samson O. Obado, Mark C. Field (Lead / Corresponding author), Brian T. Chait, Michael P. Rout (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

12 Citations (Scopus)

Abstract

Functional understanding of the nuclear envelope requires the identification of its component proteins and their interactions. Trypanosomes cause human and livestock diseases worldwide but are so divergent from animals and fungi that in silico searches for homologs of proteins are frequently of low value. Here we describe a strategy for the straightforward identification of nuclear envelope proteins from trypanosomes that classifies proteins and their interaction networks in the nuclear pore complex. Milling frozen whole cells into a powder and rapid screening of buffer conditions for optimization of complex isolation is described. The method is inexpensive and potentially applicable to many organisms, providing fast access to functional information.

Original languageEnglish
Title of host publicationThe Nuclear Envelope
EditorsSue Shackleton, Philippe Collas, Eric C. Schirmer
Place of PublicationNew York
PublisherHumana Press
Pages67-80
Number of pages14
ISBN (Electronic)9781493935307
ISBN (Print)9781493935284
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
Volume1411

Keywords

  • Nuclear envelope
  • Nuclear pore complex
  • Affinity isolation
  • Cryomilling
  • Trypanosoma
  • Proteomics
  • Molecular evolution

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