Influence of the lipid membrane environment on structure and activity of the outer membrane protein Ail from Yersinia pestis

Yi Ding, L. Miya Fujimoto, Yong Yao, Gregory V Plano, Francesca M. Marassi

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The surrounding environment has significant consequences for the structural and functional properties of membrane proteins. While native structure and function can be reconstituted in lipid bilayer membranes, the detergents used for protein solubilization are not always compatible with biological activity and, hence, not always appropriate for direct detection of ligand binding by NMR spectroscopy. Here we describe how the sample environment affects the activity of the outer membrane protein Ail (attachment invasion locus) from Yersinia pestis. Although Ail adopts the correct β-barrel fold in micelles, the high detergent concentrations required for NMR structural studies are not compatible with the ligand binding functionality of the protein. We also describe preparations of Ail embedded in phospholipid bilayer nanodiscs, optimized for NMR studies and ligand binding activity assays. Ail in nanodiscs is capable of binding its human ligand fibronectin and also yields high quality NMR spectra that reflect the proper fold. Binding activity assays, developed to be performed directly with the NMR samples, show that ligand binding involves the extracellular loops of Ail. The data show that even when detergent micelles support the protein fold, detergents can interfere with activity in subtle ways.

Original languageEnglish (US)
Pages (from-to)712-720
Number of pages9
JournalBiochimica et Biophysica Acta
Volume1848
Issue number2
DOIs
StatePublished - Feb 1 2015

Fingerprint

Yersinia pestis
Membrane Lipids
Membrane Proteins
Detergents
Ligands
Nuclear magnetic resonance
Micelles
Assays
Lipid bilayers
Lipid Bilayers
Bioactivity
Fibronectins
Nuclear magnetic resonance spectroscopy
Phospholipids
Carrier Proteins
Proteins
Magnetic Resonance Spectroscopy
Membranes

Keywords

  • Activity
  • Ail
  • Membrane protein
  • Nanodisc
  • NMR
  • Structure

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Influence of the lipid membrane environment on structure and activity of the outer membrane protein Ail from Yersinia pestis. / Ding, Yi; Fujimoto, L. Miya; Yao, Yong; Plano, Gregory V; Marassi, Francesca M.

In: Biochimica et Biophysica Acta, Vol. 1848, No. 2, 01.02.2015, p. 712-720.

Research output: Contribution to journalArticle

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