Isolation of membrane bound light-harvesting-complexes from the dinoflagellates Heterocapsa pygmaea and Prorocentrum minimum |
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Authors: | Raffael V M Jovine Geir Johnsen Barbara B Prézelin |
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Institution: | (1) Marine Primary Productivity Group, University of California at Santa Barbara, 93106 Santa Barbara, CA, USA;(2) Department of Biological Sciences, University of California, 93106 Santa Barbara, CA, USA;(3) The Marine Science Institute, University of California, 93106 Santa Barbara, CA, USA;(4) Present address: Woods Hole Oceanographic Institute, Redfield 3-32, 02543 Woods Hole, MA, USA;(5) Trondheim Biological Station, The Museum, University of Trondheim, Bynesveien 46, N-7018 Trondheim, Norway |
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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 mol m–2 s–1, HL) and low (35 mol 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.Abbreviations ACP
Chl a-Chl c-peridinin binding protein
- AEBSF
4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride
- DDM
dodecyl -d maltoside
- Deriphat 160
N-lauryl-beta-iminopropionic acid
- HEPES
(N-2-hydroxyethylpiparizine-N-2-ethanesulphonic acid)
- HL
high light (500 mol m–2 s–1)
- LL
low light (35 mol m–2 s–1)
- 730
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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Keywords: | diadinoxanthin dinoflagellate light-harvesting-complex peridinin photoacclimation photosynthesis |
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