A new approach to examine conformational changes occurring upon binding of ligand by biomolecules

Research output: Contribution to journalArticle

Abstract

Liquid-liquid partition chromatography in an aqueous poly(ethylene glycol)/dextran two-phase system (LLPC) is shown to be a quick and sensitive method for detecting conformational changes occurring upon binding of ligands by biospecific molecules. Two groups of well-characterized proteins, enzymes and monoclonal antibodies, were employed. As an example, LLPC demonstrated that isoforms of lactate dehydrogenase as well as of hexokinase existed in a ligand-dependent equilibrium between two forms and that conformational changes occurred when monoclonal antibodies bound haptens. We also demonstrate that the method could be used to detect and separate subfractions in preparations of unliganded proteins that appeared to be homogeneous when analysed by other techniques.

Details

Authors
  • Christer Wingren
  • Ulla-Britt Hansson
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biological Sciences

Keywords

  • Conformational change, Phase partition, LLPC, Ligand binding, Antibody, Enzyme
Original languageEnglish
Pages (from-to)209-215
JournalBiochimica et Biophysica Acta. General Subjects
Volume1244
Issue number1
Publication statusPublished - 1995
Publication categoryResearch
Peer-reviewedYes