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Of the vulnerability of orphan complex proteins: The case study of the E. coli IscU and IscS proteins

Prischi, Filippo and Pastore, Chiara and Carroni, Marta and Iannuzzi, Clara and Adinolfi, Salvatore and Temussi, Pierandrea and Pastore, Annalisa (2010) 'Of the vulnerability of orphan complex proteins: The case study of the E. coli IscU and IscS proteins.' Protein Expression and Purification, 73 (2). pp. 161-166. ISSN 1046-5928

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IscS and IscU, the two central protein components of the iron sulfur cluster assembly machinery, form a complex that is still relatively poorly characterized. In an attempt to standardize the purification of these proteins for structural studies we have developed a protocol to produce them individually in high concentration and purity. We show that IscS is a rather robust protein as long as it is produced in a PLP loaded form and that this co-factor is essential for fold stability and enzyme activity. In contrast to previous evidence, we also propose that, in contrast with previous evidence, IscU is a thermodynamically stable protein with a well defined fold but, when produced in isolation, is a 'complex-orphan protein' that is prone to unfolding if not stabilised by a co-factor or a protein partner. Our work will facilitate further structural and functional studies of these proteins and eventually lead to a better understanding of the whole machinery. © 2010 Elsevier Inc. All rights reserved.

Item Type: Article
Uncontrolled Keywords: Enzymatic activity; Fluorescence; Frataxin; Iron sulfur cluster; Structure
Subjects: Q Science > Q Science (General)
Divisions: Faculty of Science and Health
Faculty of Science and Health > Life Sciences, School of
SWORD Depositor: Elements
Depositing User: Elements
Date Deposited: 22 Dec 2015 16:10
Last Modified: 18 Aug 2022 11:31

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