Three-dimensional
Reconstruction of a Light-harvesting Complex I-Photosystem I (LHCI-PSI)
Supercomplex from the Green Alga Chlamydomonas reinhardtii
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adapted from Kargul J, Nield J‡, and BarberJ (2003) The Journal
of Biological Chemistry:
278(18):16135-16141
ABSTRACT:
A supercomplex containing the photosystem I (PSI)
and chlorophyll a/b light-harvesting complex I (LHCI) has been isolated
using a His-tagged mutant of Chlamydomonas reinhardtii.
This LHCI-PSI supercomplex contained .215 chlorophyll molecules of
which 175 were estimated to be chlorophyll a and 40 to be chlorophyll
b, based on P700 oxidation and chlorophyll a/b ratio measurements.
Its room temperature long wavelength absorption peak was at 680 nm,
and it emitted chlorophyll fluorescence maximally at 715 nm (77 K).
The LHCI was composed of four or more different types of Lhca polypeptides
including Lhca3. No LHCII proteins or other phosphoproteins were detected
in the LHCI-PSI supercomplexes suggesting that the cells from which
they were isolated were in State 1. Electron microscopy of negatively
stained samples followed by image analysis revealed the LHCI-PSI supercomplex
to have maximal dimensions of 220 Å by 180 Å and to be .105 Å thick.
An averaged top view was used to model in x-ray and electron crystallographic
data for PSI and Lhca proteins respectively. We conclude that the
supercomplex consists of a PSI reaction center monomer with 11 Lhca
proteins arranged along the side where the PSI proteins, PsaK, PsaJ,
PsaF, and PsaG are located. The estimated molecular mass for the complex
is 700 kDa including the bound chlorophyll molecules. The assignment
of 11 Lhca proteins is consistent with a total chlorophyll level of
215 assuming that the PSI reaction center core binds .100 chlorophylls
and that each Lhca subunit binds 10 chlorophylls. There was no evidence
for oligomerization of Chlamydomonas PSI in contrast to the
trimerization of PSI in cyanobacteria. CONTENTS
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