Immobilization of light-harvesting chlorophyll a/b complex (LHCIIb) studied by surface plasmon field-enhanced fluorescence spectroscopy.
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.
Liu
Jing
J
Max-Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
Lauterbach
Rolf
R
Paulsen
Harald
H
Knoll
Wolfgang
W
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
D045332:Photosystem II Protein Complex; D002734:Chlorophyll; D006639:Histidine; C037184:chlorophyll b; D006046:Gold; D019809:Streptavidin; D000077194:Chlorophyll A