Format

Send to

Choose Destination
Biomolecules. 2019 Sep 14;9(9). pii: E488. doi: 10.3390/biom9090488.

HDX and Native Mass Spectrometry Reveals the Different Structural Basis for Interaction of the Staphylococcal Pathogenicity Island Repressor Stl with Dimeric and Trimeric Phage dUTPases.

Author information

1
Department of Applied Biotechnology and Food Sciences, Budapest University of Technology and Economics, 1111 Budapest, Hungary. knyiri@mail.bme.hu.
2
Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary. knyiri@mail.bme.hu.
3
Department of Chemistry, King's College London, Britannia House, London SE1 1DB, UK.
4
Department of Applied Biotechnology and Food Sciences, Budapest University of Technology and Economics, 1111 Budapest, Hungary.
5
Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary.
6
Institute of Organic Chemistry, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary.
7
Department of Medical Biochemistry, Semmelweis University, 1085 Budapest, Hungary.
8
Department of Chemistry, King's College London, Britannia House, London SE1 1DB, UK. antoni.borysik@kcl.ac.uk.
9
Department of Applied Biotechnology and Food Sciences, Budapest University of Technology and Economics, 1111 Budapest, Hungary. vertessy@mail.bme.hu.
10
Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary. vertessy@mail.bme.hu.

Abstract

The dUTPase enzyme family plays an essential role in maintaining the genome integrity and are represented by two distinct classes of proteins; the β-pleated homotrimeric and the all-α homodimeric dUTPases. Representatives of both trimeric and dimeric dUTPases are encoded by Staphylococcus aureus phage genomes and have been shown to interact with the Stl repressor protein of S. aureus pathogenicity island SaPIbov1. In the present work we set out to characterize the interactions between these proteins based on a range of biochemical and biophysical methods and shed light on the binding mechanism of the dimeric φNM1 phage dUTPase and Stl. Using hydrogen deuterium exchange mass spectrometry, we also characterize the protein regions involved in the dUTPase:Stl interactions. Based on these results we provide reasonable explanation for the enzyme inhibitory effect of Stl observed in both types of complexes. Our experiments reveal that Stl employs different peptide segments and stoichiometry for the two different phage dUTPases which allows us to propose a functional plasticity of Stl. The malleable character of Stl serves as a basis for the inhibition of both dimeric and trimeric dUTPases.

KEYWORDS:

Stl staphylococcal repressor; dUTPase; inhibition; interaction surface

Supplemental Content

Full text links

Icon for Multidisciplinary Digital Publishing Institute (MDPI) Icon for PubMed Central
Loading ...
Support Center