title: Deciphering thylakoid sub-compartments using a mass spectrometry-based approach.
abstract: Photosynthesis has shaped atmospheric and ocean chemistries and probably changed the climate as well, as oxygen is released from water as part of the photosynthetic process. In photosynthetic eukaryotes, this process occurs in the chloroplast, an organelle containing the most abundant biological membrane, the thylakoids. The thylakoids of plants and some green algae are structurally inhomogeneous, consisting of two main domains: the grana, which are piles of membranes gathered by stacking forces, and the stroma-lamellae, which are unstacked thylakoids connecting the grana. The major photosynthetic complexes are unevenly distributed within these compartments because of steric and electrostatic constraints. Although proteomic analysis of thylakoids has been instrumental to define its protein components, no extensive proteomic study of subthylakoid localization of proteins in the BBY (grana) and the stroma-lamellae fractions has been achieved so far. To fill this gap, we performed a complete survey of the protein composition of these thylakoid subcompartments using thylakoid membrane fractionations. We employed semiquantitative proteomics coupled with a data analysis pipeline and manual annotation to differentiate genuine BBY and stroma-lamellae proteins from possible contaminants. About 300 thylakoid (or potentially thylakoid) proteins were shown to be enriched in either the BBY or the stroma-lamellae fractions. Overall, present findings corroborate previous observations obtained for photosynthetic proteins that used nonproteomic approaches. The originality of the present proteomic relies in the identification of photosynthetic proteins whose differential distribution in the thylakoid subcompartments might explain already observed phenomenon such as LHCII docking. Besides, from the present localization results we can suggest new molecular actors for photosynthesis-linked activities. For instance, most PsbP-like subunits being differently localized in stroma-lamellae, these proteins could be linked to the PSI-NDH complex in the context of cyclic electron flow around PSI. In addition, we could identify about a hundred new likely minor thylakoid (or chloroplast) proteins, some of them being potential regulators of the chloroplast physiology.
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; §CNRS, UMR5168, F-38054 Grenoble, France; ¶CEA, iRTSV, Laboratoire Physiologie Cellulaire & Végétale, F-38054 Grenoble, France; ‖INRA, USC 1359, F-38054 Grenoble, France;
authors.forename: Martino
authors.initials: M
authors.lastname: Tomizioli
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; **CEA, iRTSV, Laboratoire Biologie à Grande Echelle, F-38054 Grenoble, France; ‡‡ INSERM, U1038, F-38054 Grenoble, France;
authors.forename: Cosmin
authors.initials: C
authors.lastname: Lazar
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; **CEA, iRTSV, Laboratoire Biologie à Grande Echelle, F-38054 Grenoble, France; ‡‡ INSERM, U1038, F-38054 Grenoble, France;
authors.forename: Sabine
authors.initials: S
authors.lastname: Brugière
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; **CEA, iRTSV, Laboratoire Biologie à Grande Echelle, F-38054 Grenoble, France; ‡‡ INSERM, U1038, F-38054 Grenoble, France; §§CNRS, FR3425, F-38054 Grenoble, France;
authors.forename: Thomas
authors.initials: T
authors.lastname: Burger
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; §CNRS, UMR5168, F-38054 Grenoble, France; ¶CEA, iRTSV, Laboratoire Physiologie Cellulaire & Végétale, F-38054 Grenoble, France; ‖INRA, USC 1359, F-38054 Grenoble, France;
authors.forename: Daniel
authors.initials: D
authors.lastname: Salvi
authors.affiliation: ¶¶Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, CB2 1QR, United Kingdom.
authors.forename: Laurent
authors.initials: L
authors.lastname: Gatto
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; §CNRS, UMR5168, F-38054 Grenoble, France; ¶CEA, iRTSV, Laboratoire Physiologie Cellulaire & Végétale, F-38054 Grenoble, France; ‖INRA, USC 1359, F-38054 Grenoble, France;
authors.forename: Lucas
authors.initials: L
authors.lastname: Moyet
authors.affiliation: ¶¶Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, CB2 1QR, United Kingdom.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Breckels
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; **CEA, iRTSV, Laboratoire Biologie à Grande Echelle, F-38054 Grenoble, France; ‡‡ INSERM, U1038, F-38054 Grenoble, France;
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Hesse
authors.affiliation: ¶¶Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, CB2 1QR, United Kingdom.
authors.forename: Kathryn S
authors.initials: KS
authors.lastname: Lilley
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; §CNRS, UMR5168, F-38054 Grenoble, France; ¶CEA, iRTSV, Laboratoire Physiologie Cellulaire & Végétale, F-38054 Grenoble, France; ‖INRA, USC 1359, F-38054 Grenoble, France;
authors.forename: Daphné
authors.initials: D
authors.lastname: Seigneurin-Berny
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; §CNRS, UMR5168, F-38054 Grenoble, France; ¶CEA, iRTSV, Laboratoire Physiologie Cellulaire & Végétale, F-38054 Grenoble, France; ‖INRA, USC 1359, F-38054 Grenoble, France;
authors.forename: Giovanni
authors.initials: G
authors.lastname: Finazzi
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; §CNRS, UMR5168, F-38054 Grenoble, France; ¶CEA, iRTSV, Laboratoire Physiologie Cellulaire & Végétale, F-38054 Grenoble, France; ‖INRA, USC 1359, F-38054 Grenoble, France; myriam.ferro@cea.fr norbert.rolland@cea.fr.
authors.forename: Norbert
authors.initials: N
authors.lastname: Rolland
authors.affiliation: From the ‡Univ. Grenoble Alpes, F-38000 Grenoble, France; **CEA, iRTSV, Laboratoire Biologie à Grande Echelle, F-38054 Grenoble, France; ‡‡ INSERM, U1038, F-38054 Grenoble, France; myriam.ferro@cea.fr norbert.rolland@cea.fr.
authors.forename: Myriam
authors.initials: M
authors.lastname: Ferro
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D010788:Photosynthesis; D010940:Plant Proteins / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods; D020524:Thylakoids / Q000378:metabolism*
chemical_list: D010940:Plant Proteins