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The small CAB-like proteins of Synechocystis sp. PCC 6803 bind chlorophyll

In vitro pigment reconstitution studies on one-helix light-harvesting-like proteins

Abstract

The large family of light-harvesting-like proteins contains members with one to four membrane spanning helices with significant homology to the chlorophyll a/b-binding antenna proteins of plants. From structural as well as evolutionary perspective, it is likely that the members of this family bind chlorophylls and carotenoids. However, undisputable evidence is still lacking. The cyanobacterial small CAB-like proteins (SCPs) are one-helix proteins with compelling similarity to the first and third transmembrane helix of LHCII (LHCIIb) including the chlorophyll-binding motifs. They have been proposed to act as chlorophyll-carrier proteins. Here, we analyze the in vivo absorption spectra of single scp deletion mutants in Synechocystis sp. PCC 6803 and compare the in vitro pigment binding ability of the SCP pairs ScpC/D and ScpB/E with the one of LHCII and a synthetic peptide containing the chlorophyll-binding motif (Eggink LL, Hoober JK (2000) J Biol Chem 275:9087–9090). We demonstrate that deletion of scpB alters the pigmentation in the cyanobacterial cell. Furthermore, we are able to show that chlorophylls and carotenoids interact in vitro with the pairs of ScpC/D and ScpB/E, demonstrated by fluorescence resonance energy transfer and circular dichroism.

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Abbreviations

CAB:

Chlorophyll a/b binding

CD:

Circular dichroism

Chaps:

3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate

cmc:

Critical micellar concentration

DM:

n-Dodecyl-β-d-maltopyranoside

ELIP:

Early-light-induced protein

FRET:

Fluorescence resonance energy transfer

HLIP:

High-light-induced protein

LHC:

Light-harvesting complex

Lil:

Light-harvesting like

MALDI TOF MS:

Matrix assisted laser desorption ionization time of flight mass spectrometry

OG:

n-Octyl-β-d-glucopyranoside

PAGE:

Polyacrylamide gel electrophoresis

PS:

Photosystem

ROS:

Reactive oxygen species

r.t.:

Room temperature

SCP:

Small CAB-like protein

SDS:

Sodium dodecyl sulfate

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Acknowledgments

The authors would like to thank Harald Paulsen, University of Mainz, Germany, for providing the template vector for Lhcb_AB80 and for valuable discussions. The authors are also very grateful to the Royal Swedish Academy of Sciences (KVA) and to the Swedish Research Council (VR) for financial support.

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Correspondence to Christiane Funk.

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Storm, P., Hernandez-Prieto, M.A., Eggink, L.L. et al. The small CAB-like proteins of Synechocystis sp. PCC 6803 bind chlorophyll. Photosynth Res 98, 479–488 (2008). https://doi.org/10.1007/s11120-008-9368-0

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  • DOI: https://doi.org/10.1007/s11120-008-9368-0

Keywords

  • Antenna
  • Chlorophyll-binding protein
  • Cyanobacteria
  • Early-light-induced proteins (ELIPs)
  • High-light-induced proteins (HLIPs)
  • Light-harvesting complex
  • Synechocystis sp. PCC 6803