Proteomic analysis of highly purified prolamellar bodies reveals their significance in chloroplast development

• The prolamellar body (PLB) proteome of dark-grown wheat leaves was characterized. PLBs are formed not only in etioplasts but also in chloroplasts in young developing leaves during the night, yet their function is not fully understood. Highly purified PLBs were prepared from 7-day-old dark-grown leaves and identified by their spectral properties as revealed by low-temperature fluorescence spectroscopy. The PLB preparation had no contamination of extra-plastidal proteins, and only two envelope proteins were found. The PLB proteome was analysed by a combination of 1-D SDS-PAGE and nano-LC FTICR MS. The identification of chlorophyll synthase in the PLB fraction is the first time this enzyme protein was found in extracts of dark-grown plants. This finding is in agreement with its previous localization to PLBs using activity studies. NADPH:protochlorophyllide oxidoreductase A (PORA), which catalyses the reduction of protochlorophyllide to chlorophyllide, dominates the proteome of PLBs. Besides the identification of the PORA protein, the PORB protein was identified for the first time in dark-grown wheat. Altogether 64 unique proteins, representing pigment biosynthesis, photosynthetic light reaction, Calvin cycle proteins, chaperones and protein synthesis, were identified. The in number of proteins' largest group was the one involved in photosynthetic light reactions. This fact strengthens the assumption that the PLB membranes are precursors to the thylakoids and used for the formation of the photosynthetic membranes during greening. The present work is important to enhance our understanding of the significance of PLBs in chloroplast development.

Ämnesord

NATURVETENSKAP  -- Biologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences (hsv//eng)

Nyckelord

chlorophyll synthase

chloroplast development

etioplast

NADPH

protochlorophyllide oxidoreductase

prolamellar body

NADPH-PROTOCHLOROPHYLLIDE OXIDOREDUCTASE

WHEAT TRITICUM AESTIVUM

PEA

PISUM SATIVUM

ARABIDOPSIS THALIANA CHLOROPLASTS

THYLAKOID MEMBRANE

PROTEOME

II REACTION-CENTER

DARK-GROWN WHEAT

CHLOROPHYLL SYNTHETASE

ENVELOPE MEMBRANES

2-DIMENSIONAL ELECTROPHORESIS

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