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Differential proteomic analysis using iTRAQ reveals changes in thylakoids associated with Photosystem II‐acquired thermotolerance in Synechocystis sp. PCC 6803
Author(s) -
Rowland John G.,
Simon William J.,
Nishiyama Yoshitaka,
Slabas Antoni R.
Publication year - 2010
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200900337
Subject(s) - thylakoid , photosystem ii , synechocystis , biology , biochemistry , nadh dehydrogenase , photosystem i , photosynthesis , electron transport chain , cell fractionation , photosystem , biophysics , chemistry , chloroplast , enzyme , protein subunit , gene , mutant
Growth temperature has a marked influence on the thermotolerance of photosystem II (PSII), which is the most heat‐sensitive component of photosynthesis. Using Synechocystis sp. PCC 6803 we have established that thylakoids isolated from cells grown at 38°C have a greater degree of thermotolerance than those isolated from cells grown at 25°C. Reconstitution experiments using Triton X‐100 protein extracts of these thylakoids added to Triton‐treated thylakoid membranes further indicated that the 38°C Triton extract contains proteins that are directly capable of enhancing PSII thermotolerance. We have used 4‐plex iTRAQ, extensive off‐line fractionation and sample re‐injection to comprehensively identify the differences between these two preparations that may be responsible for the observed effects on PSII thermotolerance. This has resulted in the reproducible identification of 168 proteins out of a total of 385 distinct proteins. Our results have identified 15 proteins whose levels are increased in extracts that result in increased thermotolerance of PSII and 33 proteins whose levels decrease. Notably, components of the cytochrome b 6 / f and NADH dehydrogenase complexes, crucial components in electron transport, are approximately twofold more abundant in 38°C thylakoid extracts. The possible biological importance of these changes is discussed.