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Title: | Redesigning the reactive site loop of the wheat subtilisin/chymotrypsin inhibitor (WSCI) by site-directed mutagenesis. A protein–protein interaction study by affinity chromatography and molecular modeling | Authors: | Bruni, Natalia Di Maro, Antimo Costantini, Susan Chambery, Angela Facchiano, Angelo M. Ficca, Anna Grazia Parente, Augusto Poerio, Elia |
Keywords: | E/I complexes;ESI Q-TOF mass spectrometry;Time-course proteolysis;WSCI mutants | Issue Date: | 2009 | Publisher: | Elsevier | Source: | Bruni, N. et al. 2009. Redesigning the reactive site loop of the wheat subtilisin/chymotrypsin inhibitor (WSCI) by site-directed mutagenesis. A protein–protein interaction study by affinity chromatography and molecular modeling. "Biochimie". 91, 1112-1122. | Abstract: | A site-directed mutagenesis strategy was employed to obtain four mutants of wheat subtilisin/chymotrypsin inhibitor (WSCI), with the aim to produce inactive forms of this protein. The mutants were expressed in Escherichia coli as fusion proteins and, after the tag removal, were purified to homogeneity and characterized, for their interaction with the inhibited enzymes, by affinity chromatography and molecular modeling. Al fine di ottenere forme inattive di un inibitore proteico della subtilisina e della chimotripsina, è stata impiegata una strategia di mutagenesi sito-specifica. Sono stati così ottenuti quattro prodotti di espressione eterologa (mutanti dell’inibitore WSCI) che dopo purificazione sono stati sottoposti ad ampia caratterizzazione. In particolare, loro interazione con alcuni enzimi proteolitici d’interesse è stata studiata mediante cromatografia di affinità e modellamento molecolare. |
Description: | L'articolo é disponibile sul sito dell'editore: http://www.sciencedirect.com |
URI: | http://hdl.handle.net/2067/1786 | ISSN: | 0300-9084 | DOI: | 10.1016/j.biochi.2009.05.010 |
Appears in Collections: | DABAC - Archivio della produzione scientifica |
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