5C90

Staphylococcus aureus ClpP mutant - Y63A

Summary for 5C90

• Entry DOI: 10.2210/pdb5c90/pdb
• Descriptor: ATP-dependent Clp protease proteolytic subunit, (4S)-2-METHYL-2,4-PENTANEDIOL (3 entities in total)
• Functional Keywords: clp protease, proteolysis, hydrolase
• Biological source: Staphylococcus aureus
• Cellular location: Cytoplasm : Q2G036
• Total number of polymer chains: 14
• Total formula weight: 301876.51

Authors

Ye, F.,Liu, H.,Zhang, J.,Gan, J.,Yang, C.-G. (deposition date: 2015-06-26, release date: 2016-05-25, Last modification date: 2023-11-08)

Primary citation

Ni, T.,Ye, F.,Liu, X.,Zhang, J.,Liu, H.,Li, J.,Zhang, Y.,Sun, Y.,Wang, M.,Luo, C.,Jiang, H.,Lan, L.,Gan, J.,Zhang, A.,Zhou, H.,Yang, C.-G.
Characterization of Gain-of-Function Mutant Provides New Insights into ClpP Structure
Acs Chem.Biol., 11:1964-1972, 2016

PubMed Abstract: ATP-dependent Clp protease (ClpP), a highly conserved serine protease in vast bacteria, could be converted into a noncontrollable enzyme capable of degrading mature proteins in the presence of acyldepsipeptides (ADEPs). Here, we design such a gain-of-function mutant of Staphylococcus aureus ClpP (SaClpP) capable of triggering the same level of dysfunctional activity that occurs upon ADEPs treatment. The SaClpPY63A mutant degrades FtsZ in vivo and inhibits staphylococcal growth. The crystal structure of SaClpPY63A indicates that Asn42 would be an important domino to fall for further activation of ClpP. Indeed, the SaClpPN42AY63A mutant demonstrates promoted self-activated proteolysis, which is a result of an enlarged entrance pore as observed in cryo-electron microscopy images. In addition, the expression of the engineered clpP allele phenocopies treatment with ADEPs; inhibition of cell division occurs as does showing sterilizing with rifampicin antibiotics. Collectively, we show that the gain-of-function SaClpPN42AY63A mutant becomes a fairly nonspecific protease and kills persisters by degrading over 500 proteins, thus providing new insights into the structure of the ClpP protease.

PubMed: 27171654
DOI: 10.1021/acschembio.6b00390

Experimental method

X-RAY DIFFRACTION (1.75 Å)