4BTH

The LeuA146Trp,PheB24Tyr Double Mutant of the Quorum Quenching N-acyl Homoserine Lactone Acylase PvdQ Has an Altered Substrate Specificity Towards Small Acyl Chains

Summary for 4BTH

• Entry DOI: 10.2210/pdb4bth/pdb
• Related: 2WYB 2WYC 2WYD 2WYE
• Descriptor: ACYL-HOMOSERINE LACTONE ACYLASE PVDQ SUBUNIT ALPHA, ACYL-HOMOSERINE LACTONE ACYLASE PVDQ SUBUNIT BETA, GLYCEROL, ... (4 entities in total)
• Functional Keywords: zymogen, hydrolase, quorum quenching
• Biological source: PSEUDOMONAS AERUGINOSA; More
• Cellular location: Periplasm (Probable): Q9I194 Q9I194
• Total number of polymer chains: 2
• Total formula weight: 79632.36

Authors

Koch, G.,Nadal-Jimenez, P.,Reis, C.R.,Muntendam, R.,Bokhove, M.,Melillo, E.,Dijkstra, B.W.,Cool, R.H.,Quax, W.J. (deposition date: 2013-06-18, release date: 2014-01-22, Last modification date: 2024-10-16)

Primary citation

Koch, G.,Nadal-Jimenez, P.,Reis, C.R.,Muntendam, R.,Bokhove, M.,Melillo, E.,Dijkstra, B.W.,Cool, R.H.,Quax, W.J.
Reducing Virulence of the Human Pathogen Burkholderia by Altering the Substrate Specificity of the Quorum-Quenching Acylase Pvdq
Proc.Natl.Acad.Sci.USA, 111:1568-, 2014

PubMed Abstract: The use of enzymes to interfere with quorum sensing represents an attractive strategy to fight bacterial infections. We used PvdQ, an effective quorum-quenching enzyme from Pseudomonas aeruginosa, as a template to generate an acylase able to effectively hydrolyze C8-HSL, the major communication molecule produced by the Burkholderia species. We discovered that the combination of two single mutations leading to variant PvdQ(Lα146W,Fβ24Y) conferred high activity toward C8-HSL. Exogenous addition of PvdQ(Lα146W,Fβ24Y) dramatically decreased the amount of C8-HSL present in Burkholderia cenocepacia cultures and inhibited a quorum sensing-associated phenotype. The efficacy of this PvdQ variant to combat infections in vivo was further confirmed by its ability to rescue Galleria mellonella larvae upon infection, demonstrating its potential as an effective agent toward Burkholderia infections. Kinetic analysis of the enzymatic activities toward 3-oxo-C12-L-HSL and C8-L-HSL corroborated a substrate switch. This work demonstrates the effectiveness of quorum-quenching acylases as potential novel antimicrobial drugs. In addition, we demonstrate that their substrate range can be easily switched, thereby paving the way to selectively target only specific bacterial species inside a complex microbial community.

PubMed: 24474783
DOI: 10.1073/PNAS.1311263111

Experimental method

X-RAY DIFFRACTION (1.9 Å)