Purification and characterization of native human elongation factor 2

Rasmus Kock Flygaard; Beatrice Malacrida; Patrick Kiely; Lasse Bohl Jenner

doi:10.1016/j.pep.2019.02.005

Purification and characterization of native human elongation factor 2

Protein Expr Purif. 2019 Jun:158:15-19. doi: 10.1016/j.pep.2019.02.005. Epub 2019 Feb 8.

Authors

Rasmus Kock Flygaard¹, Beatrice Malacrida², Patrick Kiely², Lasse Bohl Jenner³

Affiliations

¹ Department of Molecular Biology and Genetics, University of Aarhus, Denmark.
² Graduate Entry Medical School, University of Limerick, Limerick, Ireland; Health Research Institute and Bernal Institute, University of Limerick, Limerick, Ireland.
³ Department of Integrated Structural Biology, Institute of Genetics and Molecular and Cellular Biology, CNRS UMR710, INSERM U964, University of Strasbourg, Strasbourg, 67000, France. Electronic address: lasse.jenner@igbmc.fr.

PMID: 30742898
DOI: 10.1016/j.pep.2019.02.005

Abstract

Human elongation factor 2 is the translocase that is responsible for the movement of tRNA from the A- to P- and P- to E-site on the ribosome during the elongation phase of translation. Being a vital factor of protein biosynthesis, its function is highly controlled and regulated. It has been implicated in numerous diseases and pathologies, and as such it is important to have a source for isolated pure and active protein for biomedical and biochemical studies. Here we report development of a purification protocol for native human elongation factor 2 from HEK-293S cells. The resulting protein is active, pure, has an intact diphtamide and is obtainable in yields suitable for functional and structural studies.

Keywords: Human eEF2; Translation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

HEK293 Cells
Humans
Peptide Elongation Factor 2 / chemistry*
Peptide Elongation Factor 2 / isolation & purification*

Substances

Peptide Elongation Factor 2