Smaldone, Giovanni (2013) Structural studies of proteins in intracellular environment: design and characterization of suitable cellular systems. [Tesi di dottorato]


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Item Type: Tesi di dottorato
Resource language: English
Title: Structural studies of proteins in intracellular environment: design and characterization of suitable cellular systems
Date: 28 March 2013
Number of Pages: 70
Institution: Università degli Studi di Napoli Federico II
Department: Chimica "Paolo Corradini"
Scuola di dottorato: Scienze chimiche
Dottorato: Scienze chimiche
Ciclo di dottorato: 25
Coordinatore del Corso di dottorato:
Date: 28 March 2013
Number of Pages: 70
Keywords: In cell NMR, c-prune, delivery
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/10 - Biochimica
Area 05 - Scienze biologiche > BIO/11 - Biologia molecolare
Area 03 - Scienze chimiche > CHIM/02 - Chimica fisica
Area 03 - Scienze chimiche > CHIM/09 - Farmaceutico tecnologico applicativo
Aree tematiche (7° programma Quadro): BIOTECNOLOGIE, PRODOTTI ALIMENTARI E AGRICOLTURA > Scienze della vita, biotecnologia e biochimica per prodotti e processi non-alimentari sostenibili
Date Deposited: 05 Apr 2013 05:56
Last Modified: 04 Jul 2014 11:30

Collection description

The aim of this PhD project was to design a suitable system to study a protein in its physiological conditions. This methodology becomes more intriguing when the selected molecule is an intrinsically disordered domain (IDD), whose folding may be induced by its interaction with the partners. Our target has been the IDD c-prune. Firstly we tried to obtain a fusion protein able to introduce c-prune in the cell to perform in cell-NMR spectra. To achieve this goal we have studied the mechanism of internalization of this domain in eukaryotic HeLa cell line using different CPPs. Successively the behavior of c-prune in cell lysates of eukaryotic cells has been evaluated. In particular we focused on the use of gH625 for delivery of c-prune. The peculiar properties of gH625 render it an optimal candidate to act as a carrier for net acidic charged molecules by comparison with the positively charged TAT. Furthermore this study is focused also on the interaction studies between c-prune and its different partners over-expressed in human cells via NMR spectroscopy. Besides Nm23-H1, the interaction with GSK-3beta, ASAP1 and gelsolin has also been studied. All these proteins are involved in different physiological processes that enable cells to control their shape and motility and that are strictly linked to tumor progression.


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