Assembly of G protein-coupled receptors onto nanosized bacterial magnetic particles using Mms16 as an anchor molecule

Tomoko Yoshino, Masayoshi Takahashi, Haruko Takeyama, Yoshiko Okamura, Fukuichi Kato, Tadashi Matsunaga

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

G protein-coupled receptors (GPCRs) play a central role in a wide range of biological processes and are prime targets for drug discovery. GPCRs have large hydrophobic domains, and therefore purification of GPCRs from cells is frequently time-consuming and typically results in loss of native conformation. In this work, GPCRs have been successfully assembled into the lipid membrane of nanosized bacterial magnetic particles (BMPs) produced by the magnetic bacterium Magnetospirillum magneticum AMB-1. A BMP-specific protein, Mms16, was used as an anchor molecule, and localization of heterologous Mms16 on BMPs was confirmed by luciferase fusion studies. Stable luminescence was obtained from BMPs bearing Mms16 fused with luciferase at the C-terminal region. D1 dopamine receptor (D1R), a GPCR, was also efficiently assembled onto BMPs by using Mms16 as an anchor molecule. D1R-BMP complexes were simply extracted by magnetic separation from ruptured AMB-1 transformants. After washing, the complexes were ready to use for analysis. This system conveniently refines the native conformation of GPCRs without the need for detergent solubilization, purification, and reconstitution after cell disruption.

Original languageEnglish
Pages (from-to)2880-2885
Number of pages6
JournalApplied and Environmental Microbiology
Volume70
Issue number5
DOIs
Publication statusPublished - 2004 May
Externally publishedYes

Fingerprint

G-Protein-Coupled Receptors
anchor
protein
Dopamine D1 Receptors
luciferase
Luciferases
Magnetospirillum
purification
magnetic separation
Biological Phenomena
luminescence
Drug Discovery
Membrane Lipids
Luminescence
solubilization
detergents
Detergents
washing
detergent
particle

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

Assembly of G protein-coupled receptors onto nanosized bacterial magnetic particles using Mms16 as an anchor molecule. / Yoshino, Tomoko; Takahashi, Masayoshi; Takeyama, Haruko; Okamura, Yoshiko; Kato, Fukuichi; Matsunaga, Tadashi.

In: Applied and Environmental Microbiology, Vol. 70, No. 5, 05.2004, p. 2880-2885.

Research output: Contribution to journalArticle

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