Sub-Cellular Fractionation and Gel-Based Proteomics of Haberlea Rhodopensis: A Promising Approach to Open the Black Box of Resurrection Plants

P. MLADENOV1, D. ZASHEVA2, D. PESHEV2, N. K. CHRISTOV4, N. ROLLAND5, D. DJILIANOV6 and M. TCHORBADJIEVA7
1,3,4,6 Agrobioinstitute, BG – 1164 Sofia, Bulgaria
2 Institute of Biology and Immunology of Reproduction “Acad. Kiril Bratanov“, BG – 1113 Sofia, Bulgaria
5 Institut de Recherches en Technologies et Sciences pour le Vivant (iRTSV), 17 rue des Martyrs, 38054 Grenoble cedex 9
7 Department of Biochemistry, Faculty of Biology, Sofia University “ St. Kl. Ohridski”, BG – 1164 Sofia, Bulgaria

Abstract

MLADENOV, P., D. ZASHEVA, D. PESHEV, N. K. CHRISTOV, N. ROLLAND, D. DJILIANOV and M. TCHORBADJIEVA, 2013. Sub-cellular fractionation and gel-based proteomics of Haberlea rhodopensis: a promising approach to open the black box of resurrection plants. Bulg. J. Agric. Sci., Supplement 2, 19: 22–25

 

Haberlea rhodopensis (Gesneriacea) belongs to the group of the resurrection plants. Their ability to survive extreme desiccation ren ders them excellent models to study plant response to dehydration. Plant metabolic networks are very complex and sub-cellular proteomics has proven to be a powerful approach to gain functional knowledge about the sub-cellular compartments. Here we present a protocol for the simultaneous isolation of pure and intact chloroplasts and mitochondria from Haberlea rhodopensis with subsequent gel-based proteomics including BN PAGE and 2-D gel electrophoresis. Differential detergent – based solubilization of chloroplast membranes allows separation of distinct activities in sub-organellar compartments and contributes for drastic reduction in overall sample complexity improving resolution and sensitivity of further proteomic analyses.

Key words: BN-PAGE, detergent-based subcellular fractionation, gel-based proteomics, Haberlea rhodopensis, resurrection plants
Abbreviations: 2-D gel electrophoresis – two-dimensional gel electrophoresis; SDS-PAGE – sodium dodecylsulphate polyacrylamide gel electrophoresis, BN-PAGE – Blue Native Gel Electrophoresis; DMM – dodecyl maltoside

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