Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/82682
Citations | ||
Scopus | Web of Science® | Altmetric |
---|---|---|
?
|
?
|
Type: | Journal article |
Title: | Dual roles of F123 in protein homodimerization and inhibitor binding to biotin protein ligase from Staphylococcus aureus |
Author: | Soares da Costa, T. Yap, M. Perugini, M. Wallace, J. Abell, A. Wilce, M. Polyak, S. Booker, G. |
Citation: | Molecular Microbiology, 2014; 91(1):110-120 |
Publisher: | Blackwell Publishing Ltd |
Issue Date: | 2014 |
ISSN: | 0950-382X 1365-2958 |
Statement of Responsibility: | Tatiana P. Soares da Costa, Min Y. Yap, Matthew A. Perugini, John C. Wallace, Andrew D. Abell, Matthew C. J. Wilce, Steven W. Polyak, and Grant W. Booker |
Abstract: | Protein biotinylation is catalysed by biotin protein ligase (BPL). The most characterized BPL is from Escherichia coli where it functions as both a biotin ligase and a homodimeric transcriptional repressor. Here we investigated another bifunctional BPL from the clinically important Staphylococcus aureus (SaBPL). Unliganded SaBPL (apo) exists in a dimer-monomer equilibrium at low micromolar concentrations – a stark contrast to E. coli BPL (EcBPL) that is monomeric under the same conditions. EMSA and SAXS analysis demonstrated that dimeric apo SaBPL adopted a conformation that was competent to bind DNA and necessary for it to function as a transcription factor. The SaBPL dimer-monomer dissociation constant was 5.8-fold tighter when binding the inhibitor biotin acetylene, but unchanged with biotin. F123, located in the dimer interface, was critical for homodimerization. Inhibition studies together with surface plasmon resonance analyses revealed a strong correlation between inhibitor potency and slow dissociation kinetics. A 24-fold difference in Ki values for these two enzymes was explained by differences in enzyme:inhibitor dissociation rates. Substitution of F123 in SaBPL and its equivalent in EcBPL altered both inhibitor potency and dissociation. Hence, F123 in SaBPL has novel roles in both protein dimerization and ligand-binding that have not been reported in EcBPL. |
Keywords: | Escherichia coli Staphylococcus aureus Biotin Ligases Carbon-Nitrogen Ligases Phenylalanine Bacterial Proteins Escherichia coli Proteins Repressor Proteins Ligands X-Ray Diffraction Surface Plasmon Resonance Binding Sites Amino Acid Motifs Protein Conformation Protein Structure, Quaternary Models, Molecular Scattering, Small Angle Protein Multimerization |
Rights: | © 2013 John Wiley & Sons |
DOI: | 10.1111/mmi.12446 |
Grant ID: | ARC |
Published version: | http://dx.doi.org/10.1111/mmi.12446 |
Appears in Collections: | Aurora harvest Molecular and Biomedical Science publications |
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.