Assembly of water-soluble chlorophyll-binding proteins with native hydrophobic chlorophylls in water-in-oil emulsions

Biochim Biophys Acta. 2015 Mar;1847(3):307-313. doi: 10.1016/j.bbabio.2014.12.003. Epub 2014 Dec 12.

Abstract

The challenges involved in studying cofactor binding and assembly, as well as energy- and electron transfer mechanisms in the large and elaborate transmembrane protein complexes of photosynthesis and respiration have prompted considerable interest in constructing simplified model systems based on their water-soluble protein analogs. Such analogs are also promising templates and building blocks for artificial bioinspired energy conversion systems. Yet, development is limited by the challenge of introducing the essential cofactors of natural proteins that are highly water-insoluble into the water-soluble protein analogs. Here we introduce a new efficient method based on water-in-oil emulsions for overcoming this challenge. We demonstrate the effectiveness of the method in the assembly of native chlorophylls with four recombinant variants of the water-soluble chlorophyll-binding protein of Brassicaceae plants. We use the method to gain new insights into the protein-chlorophyll assembly process, and demonstrate its potential as a fast screening system for developing novel chlorophyll-protein complexes.

Keywords: Water-in-oil emulsion; Water-soluble chlorophyll binding protein (WSCP).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brassicaceae / chemistry*
  • Brassicaceae / genetics
  • Chlorophyll / chemistry*
  • Chlorophyll / metabolism
  • Chlorophyll A
  • Chromatography, Gel
  • Emulsions
  • Hydrophobic and Hydrophilic Interactions
  • Light-Harvesting Protein Complexes / chemistry*
  • Light-Harvesting Protein Complexes / genetics
  • Light-Harvesting Protein Complexes / metabolism
  • Mineral Oil / chemistry*
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Protein Binding
  • Recombinant Proteins / chemistry
  • Solubility
  • Spectrometry, Fluorescence
  • Surface Properties
  • Water / chemistry*

Substances

  • Emulsions
  • Light-Harvesting Protein Complexes
  • Plant Proteins
  • Recombinant Proteins
  • WSCP protein, Brassica oleracea
  • Water
  • Chlorophyll
  • Mineral Oil
  • Chlorophyll A