Mutation analysis of Lhca1 antenna complex. Low energy absorption forms originate from pigment-pigment interactions.
The light harvesting complex Lhca1, one of the four gene products comprising the photosystem I antenna system, has been analyzed by site-directed mutagenesis with the aim of determining the chromophore(s) responsible for its long wavelength chlorophyll spectral form, a specific characteristic of the LHCI antenna complex. A family of mutant proteins, each carrying a mutation at a single chlorophyll-binding residue, was obtained and characterized by biochemical and spectroscopic methods. A map of the chromophores bound to each of the 10 chlorophyll-binding sites was drawn, and the energy levels of the Q(y) transition were determined in most cases. When compared with Lhcb proteins previously analyzed, Lhca1 is characterized by stronger interactions between individual chromophores as detected by both biochemical and spectroscopic methods; most mutations, although targeted to a single residue, lead to the loss of more than one chromophore and of conservative CD signals typical of chlorophyll-chlorophyll interactions. The lower energy absorption form (686 nm at 100K, 688 nm at room temperature), which is responsible for the red-shifted emission components at 690 and 701 nm, typical of Lhca1, is associated with a chlorophyll a/chlorophyll a excitonic interaction originating from a pigment cluster localized in the protein domain situated between helix C and the helix A/helix B cross. This cluster includes chlorophylls bound to sites A5-B5-B6 and a xanthophyll bound to site L2.
Morosinotto
Tomas
T
Dipartimento Scientifico e Tecnologico, Università di Verona. Strada Le Grazie, 15-37234 Verona, Italy.
Castelletti
Simona
S
Breton
Jacques
J
Bassi
Roberto
R
Croce
Roberta
R
D029681:Arabidopsis Proteins; D001665:Binding Sites; D002338:Carotenoids / Q000378:metabolism; D002734:Chlorophyll / Q000378:metabolism*; D061146:Chlorophyll Binding Proteins; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism; D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D008027:Light; D045342:Light-Harvesting Protein Complexes; D008954:Models, Biological; D009154:Mutation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics*; D045331:Photosystem I Protein Complex; D010860:Pigments, Biological / Q000378:metabolism*; D010940:Plant Proteins; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013696:Temperature; D024341:Xanthophylls / Q000737:chemistry
D029681:Arabidopsis Proteins; D061146:Chlorophyll Binding Proteins; C490278:LHCA1 protein, Arabidopsis; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D010860:Pigments, Biological; D010940:Plant Proteins; D024341:Xanthophylls; D002734:Chlorophyll; D002338:Carotenoids; D004247:DNA