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Updated: May 8, 2026
A Tandem Liquid Chromatography–Mass Spectrometry-based Approach for Metabolite Analysis of Staphylococcus aureus
Published on: March 28, 2017
Biophysical analysis of the putative acetyltransferase SACOL2570 from methicillin-resistant Staphylococcus aureus
Hai-Bin Luo1, Aleksandra A Knapik, Janusz J Petkowski
1Department of Molecular Physiology and Biological Physics, University of Virginia, 1340 Jefferson Park Avenue, Charlottesville, VA 22908, USA.
Abstract:
Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of a myriad of insidious and intractable infections in humans, especially in patients with compromised immune systems and children. Here, we report the apo- and CoA-bound crystal structures of a member of the galactoside acetyltransferase superfamily from methicillin-resistant S. aureus SACOL2570 which was recently shown to be down regulated in S. aureus grown in the presence of fusidic acid, an antibiotic used to treat MRSA infections. SACOL2570 forms a homotrimer in solution, as confirmed by small-angle X-ray scattering and dynamic light scattering. The protein subunit consists of an N-terminal alpha-helical domain connected to a C-terminal LβH domain. CoA binds in the active site formed by the residues from adjacent LβH domains. After determination of CoA-bound structure, molecular dynamics simulations were performed to model the binding of AcCoA. Binding of both AcCoA and CoA to SACOL2570 was verified by isothermal titration calorimetry. SACOL2570 most likely acts as an acetyltransferase, using AcCoA as an acetyl group donor and an as-yet-undetermined chemical moiety as an acceptor. SACOL2570 was recently used as a scaffold for mutations that lead the generation of cage-like assemblies, and has the potential to be used for the generation of more complex nanostructures.
Insights
Researchers elucidated the structure of a key protein, SACOL2570, from methicillin-resistant Staphylococcus aureus (MRSA). This protein, likely an acetyltransferase, may offer new targets for combating stubborn MRSA infections.
Area of Science:
- Biochemistry
- Structural Biology
- Microbiology
Background:
- Methicillin-resistant Staphylococcus aureus (MRSA) causes difficult-to-treat infections, particularly in vulnerable populations.
- SACOL2570, a galactoside acetyltransferase superfamily member, is downregulated by fusidic acid, an antibiotic used against MRSA.
Purpose of the Study:
- To determine the crystal structures of SACOL2570 in its apo and CoA-bound forms.
- To investigate the oligomeric state and ligand-binding properties of SACOL2570.
- To understand the potential enzymatic activity and structural applications of SACOL2570.
Main Methods:
- X-ray crystallography to determine protein structures.
- Small-angle X-ray scattering (SAXS) and dynamic light scattering (DLS) for oligomeric state analysis.
- Molecular dynamics simulations to model AcCoA binding.
- Isothermal titration calorimetry (ITC) to verify ligand binding.
Main Results:
- The apo- and CoA-bound crystal structures of SACOL2570 were determined.
- SACOL2570 exists as a homotrimer in solution.
- CoA binds within an active site formed by residues from adjacent LβH domains.
- Binding of both CoA and acetyl-CoA (AcCoA) was confirmed by ITC.
- SACOL2570 is proposed to function as an acetyltransferase.
Conclusions:
- SACOL2570's structure provides insights into its function as a potential acetyltransferase.
- The protein's ability to bind CoA and AcCoA suggests a role in Staphylococcus aureus metabolism.
- SACOL2570 serves as a scaffold for nanostructure generation, offering potential for future applications.
