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J. Biol. Chem., Vol. 277, Issue 39, 36253-36261, September 27, 2002
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From the 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 Qy 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.
Mutation Analysis of Lhca1 Antenna Complex
LOW ENERGY ABSORPTION FORMS ORIGINATE FROM PIGMENT-PIGMENT
INTERACTIONS*
,§,¶, and
Dipartimento Scientifico e Tecnologico,
Università di Verona. Strada Le Grazie, 15-37234 Verona, Italy
and the § Service de Bioénergétique,
Bâtiment 532 CEA-Saclay, 91191 Gif-sur-Yvette, France
*
This work was founded by CNR "agenzia 2000" and by
"MIUR" Progetto FIRB n.RBAU01ECX.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Present address: Istituto di biofisica, CNR Milano, c/o
Dipartimento di Biologia, Via Celoria 26, 20133 Milano, Italy.
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