Spectroscopic and molecular characterization of the oligomeric antenna of the diatom Phaeodactylum tricornutum.
The photosynthetic antenna system of diatoms contains fucoxanthin chlorophyll a/c binding proteins (FCPs), which are membrane intrinsic proteins showing high homology to the light harvesting complexes (LHC) of higher plants. In the present study, we used a mild solubilization of P. tricornutum thylakoid membranes in combination with sucrose density gradient centrifugation or gelfiltration and obtained an oligomeric FCP complex (FCPo). The spectroscopic characteristics and pigment stoichiometries of the FCPo complex were comparable to FCP complexes that were isolated after solubilization with higher detergent per chlorophyll ratios. The excitation energy transfer between the FCP-bound pigments was more efficient in the oligomeric FCPo complexes, indicating that these complexes may represent the native form of the diatom antenna system in the thylakoid membrane. Determination of the molecular masses of the two different FCP fractions by gelfiltration revealed that the FCP complexes consisted of trimers, whereas the FCPo complexes were either composed of six monomers or two tightly associated trimers. In contrast to vascular plants, stable functional monomers could not be isolated in P. tricornutum. Both types of FCP complexes showed two protein bands in SDS-gels with apparent molecular masses of 18 and 19 kDa, respectively. Sequence analysis by MS/MS revealed that the 19 kDa protein corresponded to the fcpC and fcpD genes, whereas the 18 kDa band contained the protein of the fcpE gene. The presence of an oligomeric antenna in diatoms is in line with the oligomeric organization of antenna complexes in different photoautotrophic groups.
Lepetit
Bernard
B
Institute of Biology I, Plant Physiology, University of Leipzig, Johannisallee 21-23, 04103 Leipzig, Germany.
Volke
Daniela
D
Szabó
Milán
M
Hoffmann
Ralf
R
Garab
Gyözö
G
Wilhelm
Christian
C
Goss
Reimund
R
D002734:Chlorophyll / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D017377:Diatoms / Q000378:metabolism*; D045342:Light-Harvesting Protein Complexes / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism; D020524:Thylakoids / Q000737:chemistry; D024341:Xanthophylls / Q000378:metabolism
D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D024341:Xanthophylls; C025164:fucoxanthin; D002734:Chlorophyll