Physicochemical Characterisation of KEIF-The Intrinsically Disordered N-Terminal Region of Magnesium Transporter A

Stéphanie Jephthah, Linda K. Månsson, Domagoj Belić, Jens Preben Morth, Marie Skepö*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

33 Downloads (Pure)

Abstract

Magnesium transporter A (MgtA) is an active transporter responsible for importing magnesium ions into the cytoplasm of prokaryotic cells. This study focuses on the peptide corresponding to the intrinsically disordered N-terminal region of MgtA, referred to as KEIF. Primary-structure and bioinformatic analyses were performed, followed by studies of the undisturbed single chain using a combination of techniques including small-angle X-ray scattering, circular dichroism spectroscopy, and atomistic molecular-dynamics simulations. Moreover, interactions with large unilamellar vesicles were investigated by using dynamic light scattering, laser Doppler velocimetry, cryogenic transmission electron microscopy, and circular dichroism spectroscopy. KEIF was confirmed to be intrinsically disordered in aqueous solution, although extended and containing little β -structure and possibly PPII structure. An increase of helical content was observed in organic solvent, and a similar effect was also seen in aqueous solution containing anionic vesicles. Interactions of cationic KEIF with anionic vesicles led to the hypothesis that KEIF adsorbs to the vesicle surface through electrostatic and entropic driving forces. Considering this, there is a possibility that the biological role of KEIF is to anchor MgtA in the cell membrane, although further investigation is needed to confirm this hypothesis.
Original languageEnglish
Article number623
JournalBiomolecules
Volume10
Issue number4
Number of pages22
ISSN2218-273X
DOIs
Publication statusPublished - 2020

Keywords

  • Membran proteins
  • Intrinsically disordered proteins
  • Circular dichroism spectroscopy
  • Small-angle X-ray scattering
  • Crygenic transmission electron microscopy
  • Molecular-dynamics simulations
  • Protein-vesicle interactions
  • Magnesium trnasporter
  • Secondary structure

Cite this