Engineering a two-helix bundle protein for folding studies

Charlotte Dodson, Neil Ferguson, Trevor Rutherford, Christopher Johnson, Alan Fersht

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

The SAP domain from the Saccharomyces cerevisiae THO1 protein contains a hydrophobic core and just two α-helices. It could provide a system for studying protein folding that bridges the gap between studies on isolated helices and those on larger protein domains. We have engineered the SAP domain for protein folding studies by inserting a tryptophan residue into the hydrophobic core (L31W) and solved its structure. The helical regions had a backbone root mean-squared deviation of 0.9 Å from those of wild type. The mutation L31W destabilised wild type by 0.8 ± 0.1 kcal mol−1. The mutant folded in a reversible, apparent two-state manner with a microscopic folding rate constant of around 3700 s−1 and is suitable for extended studies of folding.
Original languageEnglish
Pages (from-to)357-364
Number of pages8
JournalProtein Engineering Design and Selection
Volume23
Issue number5
Early online date3 Feb 2010
DOIs
Publication statusPublished - 1 May 2010

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