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A comparative approach towards thylakoid membrane proteome analysis of unicellular green alga Scenedesmus obliquus |
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| Authors: | Konstantinos Kantzilakis Christos Kotakis Apostolos K. Rizos Georgios Tsiotis |
| Affiliation: | a Division of Biochemistry, Department of Chemistry, University of Crete, P.O. Box 2208, 71003 Voutes Heraklion, Greece b Department of Membrane Biochemistry, Max-Planck-Institute of Biochemistry, D-82152 Martinsried, Munich, Germany c Department of Biology, University of Crete, P.O. Box 2208, GR-71409 Heraklion, Greece d Division of Physical Chemistry, Department of Chemistry, University of Crete, and Foundation for Research and Technology-Hellas (FORTH), GR-71003 Voutes Heraklion, Greece |
| Abstract: | The chlorophyll (Chl)-containing membrane protein complexes from the green alga Scenedesmus obliquus have been isolated from the thylakoid membranes by solubilization with dodecyl-β-maltoside and fractionation using a sucrose density gradient. The Chl-containing protein fractions were characterized by absorption spectroscopy, tricine SDS PAGE, BN-PAGE, and dynamic light scattering (DLS). BN-PAGE showed the presence of seven protein complexes with molecular weights in the range of 68, 118, 157, 320, 494, 828 and 955 kDa, respectively. Furthermore, light scattering reveals the simultaneous presence of particles of different sizes in the 3-4 nm and 6.0-7.5 nm range, respectively. The smaller size is related to the hydrodynamic radius of the trimer Light Harvesting Complex (LHCII), whereas the larger size is associated with the presence of photosystem I and photosystem II reaction centers. Additionally, functional information regarding protein-protein interactions was deconvoluted using coupling 2-D BN-PAGE, MALDI-TOF MS and a detailed mapping of S. obliquus photosynthetic proteome of the solubilized thylakoid membranes is therefore presented. |
| Keywords: | SDS PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis DLS, dynamic light scattering DM, Dodecyl-β-maltoside BN, blue native MALDI-TOF, matrix assisted laser desorption ionisation-time of flight LC-ESI, liquid chromatography electrospray ionisation MS, mass spectrometry |
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