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Immunological characterization of chlorophyll a/b-binding proteins of barley thylakoids

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Abstract

Monoclonal antibodies have been raised against the light-harvesting chlorophyll a/b-binding proteins of photosystem I (LHCI) using a photosystem (PS) I preparation (PSI-200) wild-type from barley (Hordeum vulgare L. cv. Svaløf's Bonus) as the antigen. These antibodies cross-reacted with a minor light-harvesting chlorophyll a/b-protein of PSII (Chla/b-P1=CP29), but not with the major one, LHCII (=Chla/b-P2**). Similarly, a monoclonal antibody to Chla/b-P1, elicited by a PSII preparation as the antigen, cross-reacted with LHCI, but not LHCII. This explains why an antigen consisting of LHCII, free of LHCI, but contaminated with Chla/b-P1, can elicit antibodies which cross-react with LHCI. Immunoblot assays showed that LHCI and Chla/b-P1 have at least two epitopes in common. Immunogold labelling of thin-sectioned wild-type thylakoids confirmed a preferential localisation of Chla/b-P1 in grana partition membranes and LHCI in stroma lamellae. The presence of LHCI was demonstrated in barley mutants lacking the PSI reaction centre (viridis-zb 63) and chlorophyll b (chlorina-f2), and was correlated with the presence of long-wavelength (730 nm) fluorescence emission at 77 K. The mutant viridis-k 23, which has a 77 K long-wavelength fluorescence peak at 720 nm, was shown by immune-blot assay to lack LHCI, although Chla/b-P1 was present.

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Abbreviations

Chl-P:

chlorophyll-protein

CM:

Carlsberg Monoclonal

Da:

dalton

LHC:

light-harvesting complex

PAGE:

polyacrylamide gel electrophoresis

PSI, II:

photosystem I, II

PSI-200:

PSI containing LHCI polypeptides

SDS:

sodium dodecyl sulphate

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Authors and Affiliations

  1. Department of Physiology, Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK-2500, Copenhagen Valby, Denmark

    G. Høyer-Hansen, L. S. Hønberg & D. J. Simpson

  2. Dipartmento di Biologia, Universitá di Padova, Via Loredan 10, I-35100, Padova, Italy

    R. Bassi

Authors
  1. G. Høyer-Hansen
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  2. R. Bassi
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  3. L. S. Hønberg
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  4. D. J. Simpson
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Høyer-Hansen, G., Bassi, R., Hønberg, L.S. et al. Immunological characterization of chlorophyll a/b-binding proteins of barley thylakoids. Planta 173, 12–21 (1988). https://doi.org/10.1007/BF00394481

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