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Chlorosome Proteins Studied by MALDI-TOF-MS: Topology of CsmA in Chlorobium tepidum

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Abstract

Chlorosomes, the light-harvesting apparatus of green bacteria, are a unique antenna system, in which pigments are organized in aggregates rather than associated with proteins. Isolated chlorosomes from the green sulphur bacterium Chlorobium tepidum contain 10 surface-exposed proteins. Treatment of chlorosomes from Chlorobium tepidum with protease caused changes in the spectral properties of bacteriochlorophyll c and digestion of chlorosome proteins. Using SDS-PAGE analysis, immunoblotting and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) we have investigated the topology of the 59-residue CsmA protein. Our results show that at the N-terminus, the only amino acid available for protease degradation is the methionine. At the C-terminus, amino acids can be removed by protease treatment to produce a residual protein containing at least the sequence between residues 2 and 38. These results indicate that the N-terminal portion of the CsmA protein, which is predicted to be mainly hydrophobic, is buried in the chlorosome envelope.

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Abbreviations

BChl:

bacteriochlorophyll

DHB:

2,5-dihydroxybenzoic acid

EDTA:

ethylenediaminetetraacetic acid

MALDI-TOF-MS:

matrix-assisted laser desorption ionization time of flight mass spectrometry

SDS:

sodium dodecyl sulfate

TFA:

trifluoroacetic acid

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Correspondence to Mette Miller.

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Milks, K.J., Danielsen, M., Persson, S. et al. Chlorosome Proteins Studied by MALDI-TOF-MS: Topology of CsmA in Chlorobium tepidum. Photosynth Res 86, 113–121 (2005). https://doi.org/10.1007/s11120-005-3757-4

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  • DOI: https://doi.org/10.1007/s11120-005-3757-4

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