Stimuli-responsive separation of proteins using immobilized liposome chromatography

Toshinori Shimanouchi, Seiichi Morita, Hiroshi Umakoshi, Ryoichi Kuboi

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The possibility of the stimuli-responsive separation of proteins was investigated using immobilized liposome chromatography (ILC) as novel aqueous two-phase systems. The specific capacity factor (k(s)) of β-galactosidase, obtained by analysis of ILC, was varied by changing the pH of the solution and was maximized at the specific pH of 5 (k(s,max) = 5.57). The k(s) values were found to correspond well with their local hydrophobicities, which can be determined by the aqueous two-phase partitioning method. The variation of k(s), therefore, indicates a change in the surface properties of a protein during conformational change under pH stimuli. A similar phenomenon is observed in the case of other proteins (α-glucosidase, k(s,max) = 11.3 at pH 4; carbonic anhydrase from bovine, k(s,max) = 6.53 at pH 4). The difference in the height and/or the position of the peaks of the k(s)-pH curves of each protein suggests a difference in their pH denaturation in the ILC column. Based on these results, the mutual separation of the above proteins at pH 4 could be successfully performed by selecting their specific capacity factor as a design parameter. (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)85-91
Number of pages7
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume743
Issue number1-2
DOIs
Publication statusPublished - Jun 23 2000
Externally publishedYes

Fingerprint

Immobilized Proteins
Chromatography
Liposomes
Proteins
Carbonic Anhydrase IV
Galactosidases
Glucosidases
Column chromatography
Denaturation
Hydrophobicity
Surface properties

Keywords

  • Immobilized liposome chromatography
  • Proteins
  • Stimuli-responsive separation

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Stimuli-responsive separation of proteins using immobilized liposome chromatography. / Shimanouchi, Toshinori; Morita, Seiichi; Umakoshi, Hiroshi; Kuboi, Ryoichi.

In: Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 743, No. 1-2, 23.06.2000, p. 85-91.

Research output: Contribution to journalArticle

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