title: Water in Oil Emulsions: A New System for Assembling Water-soluble Chlorophyll-binding Proteins with Hydrophobic Pigments.
abstract: Chlorophylls (Chls) and bacteriochlorophylls (BChls) are the primary cofactors that carry out photosynthetic light harvesting and electron transport. Their functionality critically depends on their specific organization within large and elaborate multisubunit transmembrane protein complexes. In order to understand at the molecular level how these complexes facilitate solar energy conversion, it is essential to understand protein-pigment, and pigment-pigment interactions, and their effect on excited dynamics. One way of gaining such understanding is by constructing and studying complexes of Chls with simple water-soluble recombinant proteins. However, incorporating the lipophilic Chls and BChls into water-soluble proteins is difficult. Moreover, there is no general method, which could be used for assembly of water-soluble proteins with hydrophobic pigments. Here, we demonstrate a simple and high throughput system based on water-in-oil emulsions, which enables assembly of water-soluble proteins with hydrophobic Chls. The new method was validated by assembling recombinant versions of the water-soluble chlorophyll binding protein of Brassicaceae plants (WSCP) with Chl a. We demonstrate the successful assembly of Chl a using crude lysates of WSCP expressing E. coli cell, which may be used for developing a genetic screen system for novel water-soluble Chl-binding proteins, and for studies of Chl-protein interactions and assembly processes.
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science.
authors.forename: Dominika
authors.initials: D
authors.lastname: Bednarczyk
authors.affiliation: Migal-Galilee Research Institute; drorn@migal.org.il.
authors.forename: Dror
authors.initials: D
authors.lastname: Noy
mesh_terms: D001937:Brassica / Q000737:chemistry; D002734:Chlorophyll / Q000737:chemistry; D061146:Chlorophyll Binding Proteins / Q000737:chemistry* / Q000235:genetics; D002942:Circular Dichroism; D004655:Emulsions; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D057927:Hydrophobic and Hydrophilic Interactions; D045342:Light-Harvesting Protein Complexes / Q000737:chemistry*; D009821:Oils / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D014867:Water / Q000737:chemistry*
chemical_list: D061146:Chlorophyll Binding Proteins; D004655:Emulsions; D045342:Light-Harvesting Protein Complexes; D009821:Oils; D011994:Recombinant Proteins; D014867:Water; D002734:Chlorophyll