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Isolation of membrane bound light-harvesting-complexes from the dinoflagellates Heterocapsa pygmaea and Prorocentrum minimum

  • Light-Harvesting Complexes
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Abstract

We have isolated Chl a-Chl c-carotenoid binding proteins from the dinoflagellates Prorocentrum minimum and Heterocapsa pygmaea grown under high (500 μmol m−2 s−1, HL) and low (35 μmol m−2 s−1, LL) light conditions. We compared various isolation procedures of membrane bound light harvesting complexes (LHCs) and assayed the functionality of the solubilized proteins by determining the energy transfer efficiency from the accessory pigments to Chl a by means of fluorescence excitation spectra. The identity of the newly isolated protein-complexes were confirmed by immunological cross-reactions with antibodies raised against the previously described membrane bound Chl a-c proteins (Boczar et al. (1980) FEBS Lett 120: 243–247). Spectroscopic analysis demonstrated the relatedness of these proteins with the recently described Chl-a-c 2-peridinin (ACP) binding protein (Hiller et al. (1993) Photochem Photobiol 57: 125–131; Iglesias Prieto et al. (1993) Phil Trans R Soc London B 338: 381–392). The water-soluble peridinin-Chl a binding-protein (PCP) was not detectable in P. minimum. Two functional forms of ACP with different pigmentation were isolated. A variant of ACP which was isolated from high-light grown cells, that specifically binds increased amounts of diadinoxanthin was compared to the previously described ACPs that bind proportionately more peridinin.

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Abbreviations

ACP:

Chl a-Chl c-peridinin binding protein

AEBSF:

4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride

DDM:

dodecyl β-d maltoside

Deriphat 160:

N-lauryl-beta-iminopropionic acid

HEPES:

(N-2-hydroxyethylpiparizine-N′-2-ethanesulphonic acid)

HL:

high light (500 μmol m−2 s−1)

LL:

low light (35 μmol m−2 s−1)

Ф730 :

fluorescence yield (emission at 730 nm)

PCP:

peridinin-Chl a-binding protein

PMSF:

phenyl-methyl-sulfonyl-fluoride

PS I:

Photosystem I

PS II:

Photosystem II

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Author notes

  1. Raffael V. M. Jovine

    Present address: Woods Hole Oceanographic Institute, Redfield 3-32, 02543, Woods Hole, MA, USA

Authors and Affiliations

  1. Marine Primary Productivity Group, University of California at Santa Barbara, 93106, Santa Barbara, CA, USA

    Raffael V. M. Jovine, Geir Johnsen & Barbara B. Prézelin

  2. Department of Biological Sciences, University of California, 93106, Santa Barbara, CA, USA

    Raffael V. M. Jovine, Geir Johnsen & Barbara B. Prézelin

  3. The Marine Science Institute, University of California, 93106, Santa Barbara, CA, USA

    Raffael V. M. Jovine, Geir Johnsen & Barbara B. Prézelin

  4. Trondheim Biological Station, The Museum, University of Trondheim, Bynesveien 46, N-7018, Trondheim, Norway

    Geir Johnsen

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Jovine, R.V.M., Johnsen, G. & Prézelin, B.B. Isolation of membrane bound light-harvesting-complexes from the dinoflagellates Heterocapsa pygmaea and Prorocentrum minimum . Photosynth Res 44, 127–138 (1995). https://doi.org/10.1007/BF00018303

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