Structural analysis of Escherichia coli ThiF

Duda, D. M., Walden, H. , Sfondouris, J. and Schulman, B. A. (2005) Structural analysis of Escherichia coli ThiF. Journal of Molecular Biology, 349(4), pp. 774-786. (doi: 10.1016/j.jmb.2005.04.011) (PMID:15896804)

Full text not currently available from Enlighten.


Escherichia coli ThiF is an enzyme in the biosynthetic cascade for generating the essential cofactor thiamin pyrophosphate. In this cascade, ThiF catalyzes adenylation of the C terminus of ThiS. We report here the crystal structures of ThiF, alone and in complex with ATP. The structures provide insight into a preference for ATP during adenylation of the protein ThiS. Additionally, the structures reveal an ordered crossover loop predicted to clamp the flexible tail of ThiS into the ThiF active site during the adenylation reaction. The importance of the crossover loop for ThiF activity is highlighted by mutational analysis. Comparison of ThiF with the structural homologues MoeB, APPBP1-UBA3, and SAE1-SAE2 reveals that the ATP-binding site, including an arginine-finger, is maintained throughout evolution, and shows divergence occurring in protein substrate-binding sites and regions devoted to unique steps in the specific function of each enzyme.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Walden, Professor Helen
Authors: Duda, D. M., Walden, H., Sfondouris, J., and Schulman, B. A.
College/School:College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
Journal Name:Journal of Molecular Biology
ISSN (Online):1089-8638
Published Online:04 May 2005

University Staff: Request a correction | Enlighten Editors: Update this record