3V5I

The crystal structure of the mutant ClpP S98A (Staphylococcus aureus)

Summary for 3V5I

• Entry DOI: 10.2210/pdb3v5i/pdb
• Related: 3QWD 3V5E
• Descriptor: ATP-dependent Clp protease proteolytic subunit (2 entities in total)
• Functional Keywords: hydrolase, whole proteome
• Biological source: Staphylococcus aureus subsp. aureus
• Cellular location: Cytoplasm : Q2G036
• Total number of polymer chains: 28
• Total formula weight: 631755.04

Authors

List, A.,Gersch, M.,Groll, M.,Sieber, S. (deposition date: 2011-12-16, release date: 2012-02-08, Last modification date: 2023-09-13)

Primary citation

Gersch, M.,List, A.,Groll, M.,Sieber, S.A.
Insights into structural network responsible for oligomerization and activity of bacterial virulence regulator caseinolytic protease P (ClpP) protein.
J.Biol.Chem., 287:9484-9494, 2012

PubMed Abstract: The barrel-shaped caseinolytic protease P (ClpP) is a main virulence regulator in the bacterial pathogen Staphylococcus aureus (SaClpP). It consists of two heptameric rings forming a homotetradecamer with an inner chamber that houses the 14 active sites. We recently showed that SaClpP is able to adopt a compressed, inactive conformation. We present here the 2.3 Å resolution structure of SaClpP in its closed, active conformation as well as the structure of the S98A mutant. Comprehensive mutational analysis aiming at destabilizing one or the other or both conformations was able to pinpoint key residues involved in this catalytic switch and in the heptamer-heptamer interaction. By probing the active site serine with a covalently modifying β-lactone probe, we could show that the tetradecameric organization is essential for a proper formation of the active site. Structural data suggest that a highly conserved hydrogen-bonding network links oligomerization to activity. A comparison of ClpP structures from different organisms provides suggestive evidence for the presence of a universal mechanism regulating ClpP activity in which binding of one subunit to the corresponding subunit on the other ring interface is necessary for the functional assembly of the catalytic triad and thus for protease function. This mechanism ensures controlled access to the active sites of a highly unspecific protease.

PubMed: 22291011
DOI: 10.1074/jbc.M111.336222

Experimental method

X-RAY DIFFRACTION (2.8 Å)