title: Immobilization of light-harvesting chlorophyll a/b complex (LHCIIb) studied by surface plasmon field-enhanced fluorescence spectroscopy.
abstract: The major light-harvesting chlorophyll a/ b complex (LHCIIb) of the photosynthetic apparatus in green plants can be viewed as a protein scaffold binding and positioning a large number of pigment molecules that engage in rapid excitation energy transfer. This property makes LHCIIb potentially interesting as a light harvester (or a model thereof) in photoelectronic applications. Such applications would require the immobilization of LHCIIb (or similar dye-protein complexes) on a solid surface. In this work, the immobilization of recombinant LHCIIb is tested and optimized on functionalized gold surfaces via a histidine 6 tag (His tag) in the protein moiety. Immobilization efficiency and kinetics are analyzed by using surface plasmon resonance (SPR) and surface plasmon field-enhanced fluorescence spectroscopy (SPFS). The latter was also used to assess the integrity of immobilized LHCIIb by recording Chl b-sensitized Chl a emission spectra. Since His tags have been included in a substantial number of recombinant proteins, the immobilization technique developed here for LHCIIb presumably can be extended to a large range of other membrane and water-soluble proteins.
authors.affiliation: Max-Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Rolf
authors.initials: R
authors.lastname: Lauterbach
authors.forename: Harald
authors.initials: H
authors.lastname: Paulsen
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Knoll
mesh_terms: D019921:Biotinylation; D002734:Chlorophyll / Q000737:chemistry*; D000077194:Chlorophyll A; D006046:Gold; D006639:Histidine / Q000737:chemistry; D007700:Kinetics; D008027:Light; D008956:Models, Chemical; D008968:Molecular Conformation; D010777:Photochemistry / Q000379:methods; D045332:Photosystem II Protein Complex / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D019809:Streptavidin / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000379:methods*; D013997:Time Factors
chemical_list: D045332:Photosystem II Protein Complex; D002734:Chlorophyll; D006639:Histidine; C037184:chlorophyll b; D006046:Gold; D019809:Streptavidin; D000077194:Chlorophyll A