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Proteomic profiles of thylakoid membranes and changes in response to iron deficiency

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Abstract

The proteomic profile of thylakoid membranes and the changes induced in that proteome by iron deficiency have been studied by using thylakoid preparations from Beta vulgaris plants grown in hydroponics. Two different 2-D electrophoresis approaches have been used to study these proteomes: isoelectrical focusing followed by SDS PAGE (IEF-SDS PAGE) and blue-native polyacrylamide gel electrophoresis followed by SDS PAGE (BN-SDS PAGE). These techniques resolved approximately 110–140 and 40 polypeptides, respectively. Iron deficiency induced significant changes in the thylakoid sugar beet proteome profiles: the relative amounts of electron transfer protein complexes were reduced, whereas those of proteins participating in leaf carbon fixation-linked reactions were increased. A set of polypeptides, which includes several enzymes related to metabolism, was detected in thylakoid preparations from Fe-deficient Beta vulgaris leaves by using BN-SDS PAGE, suggesting that they may be associated with these thylakoids in vivo. The BN-SDS PAGE technique has been proven to be a better method than IEF-SDS PAGE to resolve highly hydrophobic integral membrane proteins from thylakoid preparations, allowing for the identification of complexes and determination of their polypeptidic components.

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Abbreviations

IEF-SDS PAGE:

Isoelectrical focusing polyacrylamide gel electrophoresis

BN-SDS PAGE:

Blue-native polyacrylamide gel electrophoresis

LC-MS:

Liquid chromatography mass spectrometry

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Acknowledgements

This work was supported by Spanish Ministry of Science and Education (MEC) projects BOS2001-2343, AGL2003-1999 and AGL2004-0194 (co-financed with FEDER). SA was supported by a predoctoral fellowship from the MEC. AF L-M was scientist on a “Ramón y Cajal” contract from the Spanish Ministry of Education and Culture. We thank Aurora Poc (Estación Experimental de Aula Dei) for excellent technical assistance in growing the plants, and Virpi Paakkarinen and Sari Sirpiö (University of Turku) for their precious help in BN techniques.

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Authors and Affiliations

  1. Department of Plant Nutrition, Estación Experimental de Aula Dei, CSIC, Zaragoza, Spain

    Sofía Andaluz, Ana-Flor López-Millán, Javier Abadía & Anunciación Abadía

  2. Bioinformatics and Functional Genomics Research Group, Cancer Research Center (CIC), University of Salamanca & CSIC, Salamanca, Spain

    Javier De las Rivas

  3. Plant Physiology and Molecular Biology, Department of Biology, University of Turku, Turku, Finland

    Eva-Mari Aro

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  1. Sofía Andaluz
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  2. Ana-Flor López-Millán
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Correspondence to Anunciación Abadía.

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Andaluz, S., López-Millán, AF., De las Rivas, J. et al. Proteomic profiles of thylakoid membranes and changes in response to iron deficiency. Photosynth Res 89, 141–155 (2006). https://doi.org/10.1007/s11120-006-9092-6

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