Evaluation of temperature and guanidine hydrochloride-induced protein-liposome interactions by using immobilized liposome chromatography

Noriko Yoshimoto, Makoto Yoshimoto, Kazuma Yasuhara, Toshinori Shimanouchi, Hiroshi Umakoshi, Ryoichi Kuboi

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Hydrophobic interactions between nine model proteins and net-neutral lipid bilayer membranes (liposomes) under stress conditions were quantitatively examined by using immobilized liposome chromatography (ILC). Small or large unilamellar liposomes were composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and immobilized in a gel matrix by utilizing covalent coupling between amino-containing lipids and activated gel beads or avidin-biotin biospecific binding. Retardation of bovine carbonic anhydrase (CAB) in ILC was pronounced at particular temperatures (50 and 60 °C) where the local hydrophobicity of theses protein molecules becomes sufficiently large. Protein-induced leakage of a hydrophilic dye (calcein) from immobilized liposomes interior was also drastically enhanced at particular temperatures where large retardation was observed. For other proteins examined, similar results were also observed. The specific capacity factor of the proteins characteristic for the ILC and the amount of calcein released from immobilized liposomes were successfully expressed as a function of the product of the local hydrophobicities of proteins and liposomes, regardless of protein species and the type of the stress conditions applied (denaturant and heating). These findings indicate that lipid membranes have an ability to non-specifically recognize local hydrophobicities of proteins to form stress-mediated supramolecular assemblies with proteins, which may have potential applications in bioprocesses such as protein refolding and separation. ILC was thus found to be a very useful method for the quantitative detection of dynamic protein-liposome interactions triggered by stress conditions.

Original languageEnglish
Pages (from-to)174-181
Number of pages8
JournalBiochemical Engineering Journal
Volume29
Issue number3
DOIs
Publication statusPublished - Apr 15 2006
Externally publishedYes

Keywords

  • Biosensors
  • Bioseperations
  • Immobilized liposome chromatography
  • Membrane stress biotechnology
  • Protein
  • Protein denaturation

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Engineering
  • Bioengineering
  • Biomedical Engineering

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