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Anisotropic circular dichroism signatures of oriented thylakoid membranes and lamellar aggregates of LHCII

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Abstract

In photosynthesis research, circular dichroism (CD) spectroscopy is an indispensable tool to probe molecular architecture at virtually all levels of structural complexity. At the molecular level, the chirality of the molecule results in intrinsic CD; pigment–pigment interactions in protein complexes and small aggregates can give rise to excitonic CD bands, while “psi-type” CD signals originate from large, densely packed chiral aggregates. It has been well established that anisotropic CD (ACD), measured on samples with defined non-random orientation relative to the propagation of the measuring beam, carries specific information on the architecture of molecules or molecular macroassemblies. However, ACD is usually combined with linear dichroism and can be distorted by instrumental imperfections, which given the strong anisotropic nature of photosynthetic membranes and complexes, might be the reason why ACD is rarely studied in photosynthesis research. In this study, we present ACD spectra, corrected for linear dichroism, of isolated intact thylakoid membranes of granal chloroplasts, washed unstacked thylakoid membranes, photosystem II (PSII) membranes (BBY particles), grana patches, and tightly stacked lamellar macroaggregates of the main light-harvesting complex of PSII (LHCII). We show that the ACD spectra of face- and edge-aligned stacked thylakoid membranes and LHCII lamellae exhibit profound differences in their psi-type CD bands. Marked differences are also seen in the excitonic CD of BBY and washed thylakoid membranes. Magnetic CD (MCD) spectra on random and aligned samples, and the largely invariable nature of the MCD spectra, despite dramatic variations in the measured isotropic and anisotropic CD, testify that ACD can be measured without substantial distortions and thus employed to extract detailed information on the (supra)molecular organization of photosynthetic complexes. An example is provided showing the ability of CD data to indicate such an organization, leading to the discovery of a novel crystalline structure in macroaggregates of LHCII.

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Abbreviations

ACD:

Anisotropic circular dichroism

BBY:

Photosystem II-enriched grana membrane fragments, isolated according to Berthold, Babcock and Yocum

CD:

Circular dichroism

CDexc :

Excitonic circular dichroism

CDψ :

Psi-type circular dichroism

Chl:

Chlorophyll

LD:

Linear dichroism

LHCII:

Light-harvesting complex of Photosystem II

MCD:

Magnetic circular dichroism

PSI, PSII:

Photosystem I, II

STEM:

Scanning transmission electron microscopy

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Acknowledgments

This study was supported by the EC Marie Curie Training Network (ITN) “HARVEST” (No. 238017) and the Hungarian Scientific Research Fund, OTKA/NKTH CNK 80345 to G.G. and GOP-1.1.2-07/1-2008-0007 to P.L. The authors thank Hans-Georg Kuball for his helpful advice, László Kovács for the preparation of BBY and LHCII, Beth Lin for the STEM sample preparation and Martha Simon for the STEM operation. US-NIH and US-DOE provided financial support to the STEM Facility. GH received support from US-DOE, Office of Basic Energy Sciences, Division of Energy Biosciences.

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Authors and Affiliations

  1. Institute of Plant Biology, Biological Research Center, Hungarian Academy of Sciences, P.O. Box 521, 6701, Szeged, Hungary

    Yuliya Miloslavina, Petar H. Lambrev, Zsuzsanna Várkonyi, Václav Karlický & Győző Garab

  2. Diamond Light Source Ltd., Diamond House, Harwell Science and Innovation Campus, Chilton, Didcot, OX11 0DE, UK

    Tamás Jávorfi

  3. Biology Department, Brookhaven National Laboratory, Upton, NY, USA

    Joseph S. Wall & Geoffrey Hind

Authors
  1. Yuliya Miloslavina
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  2. Petar H. Lambrev
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  3. Tamás Jávorfi
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  4. Zsuzsanna Várkonyi
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  5. Václav Karlický
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  7. Geoffrey Hind
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  8. Győző Garab
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Correspondence to Győző Garab.

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Miloslavina, Y., Lambrev, P.H., Jávorfi, T. et al. Anisotropic circular dichroism signatures of oriented thylakoid membranes and lamellar aggregates of LHCII. Photosynth Res 111, 29–39 (2012). https://doi.org/10.1007/s11120-011-9664-y

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