Bead-linked proteoliposomes: A reconstitution method for NMR analyses of membrane protein-ligand interactions

Mariko Yokogawa, Koh Takeuchi, Ichio Shimada

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Structural information about the interactions between membrane proteins and their ligands provides insights into the membrane protein functions. A variety of surfactants have been used for structural analyses of membrane proteins, and in some cases, they yielded successful results. However, the use of surfactants frequently increases the conformational instability of membrane proteins and distorts their normal function. Here, we propose a new strategy of membrane protein reconstitution into lipid bilayers on affinity beads, which maintains the native conformation and function of the protein for NMR studies. The reconstituted membrane proteins are suitable for NMR analyses of interactions, by using the transferred cross-saturation method. The strategy was successfully applied to the interaction between a potassium ion channel, KcsA, and a pore-blocker, agitoxin2 (AgTx2). This strategy would be useful for analyzing the interactions between various membrane proteins and their ligands.

Original languageEnglish
Pages (from-to)12021-12027
Number of pages7
JournalJournal of the American Chemical Society
Volume127
Issue number34
DOIs
Publication statusPublished - 2005 Aug 31
Externally publishedYes

    Fingerprint

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this