Towards an understanding of wheat chloroplasts: a methodical investigation of thylakoid proteome |
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Authors: | Abu?Hena?Mostafa?Kamal Kun?Cho Setsuko?Komatsu Nobuyuki?Uozumi Jong-Soon?Choi Email author" target="_blank">Sun?Hee?WooEmail author |
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Institution: | Department of Crop Science, Chungbuk National University, 410 Seongbong-ro, Heungdeok-gu, Cheongju, Chungbuk, 361-763, Korea. |
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Abstract: | We utilized Percoll density gradient centrifugation to isolate and fractionate chloroplasts of Korean winter wheat cultivar
cv. Kumgang (Triticum aestivum L.). The resulting protein fractions were separated by one dimensional polyacrylamide gel electrophoresis (1D-PAGE) coupled
with LTQ-FTICR mass spectrometry. This enabled us to detect and identify 767 unique proteins. Our findings represent the most
comprehensive exploration of a proteome to date. Based on annotation information from the UniProtKB/Swiss-Prot database and
our analyses via WoLF PSORT and PSORT, these proteins are localized in the chloroplast (607 proteins), chloroplast stroma
(145), thylakoid membrane (342), lumens (163), and integral membranes (166). In all, 67% were confirmed as chloroplast thylakoid
proteins. Although nearly complete protein coverage (89% proteins) has been accomplished for the key chloroplast pathways
in wheat, such as for photosynthesis, many other proteins are involved in regulating carbon metabolism. The identified proteins
were assigned to 103 functional categories according to a classification system developed by the iProClass database and provided through Protein Information Resources. Those functions include electron transport, energy,
cellular organization and biogenesis, transport, stress responses, and other metabolic processes. Whereas most of these proteins
are associated with known complexes and metabolic pathways, about 13% of the proteins have unknown functions. The chloroplast
proteome contains many proteins that are localized to the thylakoids but as yet have no known function. We propose that some
of these familiar proteins participate in the photosynthetic pathway. Thus, our new and comprehensive protein profile may
provide clues for better understanding that photosynthetic process in wheat. |
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