http://www.cnr.it/ontology/cnr/individuo/prodotto/ID8957
Molecular cloning, expression and characterization of a novel apoplastic invertase inhibitor from tomato (Solanum lycopersicum) and its use to purify a vacuolar invertase. (Articolo in rivista)
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- Molecular cloning, expression and characterization of a novel apoplastic invertase inhibitor from tomato (Solanum lycopersicum) and its use to purify a vacuolar invertase. (Articolo in rivista) (literal)
- Anno
- 2008-01-01T00:00:00+01:00 (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi
- 10.1016/j.biochi.2008.04.019 (literal)
- Alternative label
Ida Barbara Reca ; Alexandre Brutus ; Rossana D'Avino ; Claude Villard ; Daniela Bellincampi b, Thierry Giardina (2008)
Molecular cloning, expression and characterization of a novel apoplastic invertase inhibitor from tomato (Solanum lycopersicum) and its use to purify a vacuolar invertase.
in Biochimie (Print); ELSEVIER SCIENCE BV, PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS, AMSTERDAM (Paesi Bassi)
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- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
- Ida Barbara Reca ; Alexandre Brutus ; Rossana D'Avino ; Claude Villard ; Daniela Bellincampi b, Thierry Giardina (literal)
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- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#url
- http://www.sciencedirect.com/science/article/pii/S0300908408001843 (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume
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- ISI Web of Science (WOS) (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
- ISM2/BiosCiences UMR CNRS 6263 (Universite´ Aix Marseille III/CNRS), Inge´nierie et Me´canismes d'Action des Glycosidases, service 342 Faculte´ des Sciences et Techniques Saint-Je´ro^me, Universite´ Paul Ce´zanne, 13397 Marseille Cedex 20, France ; Dipartimento di Biologia Vegetale, Universita` di Roma La Sapienza, Piazzale Aldo Moro 5, Roma 00185, Italy ; Institute of Protein Biochemistry, CNR, Via P. Castellino 111, 80131 Naples, Italy ; Plateau prote´omique UMR FRE 2737, Laboratoire de Nutrition & Die´te´tique, Faculte´ de pharmacie, Universite´ de Me´diterrane´e,27 av. J. Moulin, 13385 Marseille Cedex 5, France (literal)
- Titolo
- Molecular cloning, expression and characterization of a novel apoplastic invertase inhibitor from tomato (Solanum lycopersicum) and its use to purify a vacuolar invertase. (literal)
- Abstract
- Protein inhibitors are molecules secreted by many plants. In a functional genomics approach, an invertase inhibitor (SolyCIF) of Solanum lycopersicum was identified at the Solanaceae Cornell University data bank (www.sgn.cornell.edu). It was established that this inhibitor is expressed mainly in the leaves, flowers and green fruit of the plant and localized in the cell wall compartment. The SolyCIF cDNA was cloned by performing RT-PCR, fully sequenced and heterologously expressed in Pichia pastoris X-33. The purified recombinant protein obtained by performing ion-exchange chromatography and gel filtration was further biochemically characterized and used to perform affinity chromatography. The latter step made it possible to purify natural vacuolar invertase (TIV-1), which showed high rates of catalytic activity (438.3 U.mg-1) and efficiently degraded saccharose (Km = 6.4 mM, Vmax=2.9µmol saccharose min-1 and kcat = 7.25 103sec-1 at pH 4.9 and 37° C). The invertase activity was strongly inhibited in a dose-dependent manner by SolyCIF produced in P. pastoris. In addition, Gel-SDS PAGE analysis strongly suggest that TIV-1 was proteolyzed in planta and it was established that the fragments produced have to be tightly associated for its enzymatic activity to occur. We further investigated the location of the proteolytic sites by performing NH2-terminal Edman degradation on the fragments. The molecular model for TIV-1 shows that the fragmentation splits the catalytic site of the enzyme into two halves, which confirms that the enzymatic activity is possible only when the fragments are tightly associated. (literal)
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