Stable oil bodies sheltered by a unique caleosin in cycad megagametophytes |
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Authors: | Pei-Luen Jiang Jeff CF Chen Shau-Ting Chiu Jason TC Tzen |
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Institution: | 1. Jiangsu Key Laboratory for Molecular & Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing 210023, PR China;2. Basic Medical College, Nanjing University of Chinese Medicine, Nanjing 210023, PR China;3. Jiangsu Key Laboratory for Aquatic Crustacean Diseases, Life Science College, Nanjing Normal University, Nanjing 210023, PR China;4. Jiangsu Key Laboratory for Supramolecular Medicinal Materials & Applications, Life Science College, Nanjing Normal University, Nanjing 210023, PR China;1. INRA, Institut Jean-Pierre Bourgin, UMR 1318, ERL CNRS 3559, Saclay Plant Sciences, RD10, F-78026 Versailles, France;2. AgroParisTech, Institut Jean-Pierre Bourgin, UMR 1318, ERL CNRS 3559, Saclay Plant Sciences, RD10, F-78026 Versailles, France |
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Abstract: | Stable oil bodies of smaller sizes and higher thermostability were isolated from mature cycad (Cycas revoluta) megagametophytes compared with those isolated from sesame seeds. Immunological cross-recognition revealed that cycad oil bodies contained a major protein of 27 kDa, tentatively identified as caleosin, while oleosin, the well-known structural protein, was apparently absent. Mass spectrometric analysis showed that the putative cycad caleosin possessed a tryptic fragment of 15 residues matching to that of a theoretical moss caleosin. A complete cDNA fragment encoding this putative caleosin was obtained by PCR cloning using a primer designed according to the tryptic peptide and another one designed according to a highly conservative region among diverse caleosins. The identification of this clone was subsequently confirmed by immunodetection and MALDI-MS analyses of its recombinant fusion protein over-expressed in Escherichia coli and the native form from cycad oil bodies. Stable artificial oil bodies were successfully constituted with triacylglycerol, phospholipid and the recombinant fusion protein containing the cycad caleosin. These results suggest that stable oil bodies in cycad megagametophytes are mainly sheltered by a unique structural protein caleosin. |
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