Details
| Original language | English |
|---|---|
| Article number | e202300185 |
| Number of pages | 7 |
| Journal | CHEMBIOCHEM |
| Volume | 24 |
| Issue number | 17 |
| Early online date | 17 May 2023 |
| Publication status | Published - 1 Sept 2023 |
Abstract
The human pathogen Pseudomonas aeruginosa has a number of virulence factors at its disposal that play crucial roles in the progression of infection. LasB is one of the major virulence factors and exerts its effects through elastolytic and proteolytic activities aimed at dissolving connective tissue and inactivating host defense proteins. LasB is of great interest for the development of novel pathoblockers to temper the virulence, but access has thus far largely been limited to protein isolated from Pseudomonas cultures. Here, we describe a new protocol for high-level production of native LasB in Escherichia coli. We demonstrate that this facile approach is suitable for the production of mutant, thus far inaccessible LasB variants, and characterize the proteins biochemically and structurally. We expect that easy access to LasB will accelerate the development of inhibitors for this important virulence factor.
Keywords
- anti-infective agents, LasB, pathoblockers, virulence factors
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Medicine
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Chemistry(all)
- Organic Chemistry
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In: CHEMBIOCHEM, Vol. 24, No. 17, e202300185, 01.09.2023.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Facile Production of the Pseudomonas aeruginosa Virulence Factor LasB in Escherichia coli for Structure-Based Drug Design
AU - Kolling, Dominik
AU - Haupenthal, Jörg
AU - Hirsch, Anna K.H.
AU - Koehnke, Jesko
N1 - Funding Information: 1 . We acknowledge the use of the ESRF beamlines ID30B and ID23‐1. We thank Simone Amann for the PA cultivation and supernatant preparation and Timo Risch for conducting the protein LC–MS measurements. We thank Dr. Andreas Kany for kindly providing compound . We thank Dr. Sally Shirran from the mass spectrometry and proteomics facility at St. Andrews University for conducting the tryptic LasB digesting and MS/MS measurements. Figure 1 2 and the table of contents graphic were prepared by using BioRender.com (2023). This work was supported by an ANR/BMBF grant (LasBAntiv, 16GW0346) to A.K.H.H. and J.K. Open Access funding enabled and organized by Projekt DEAL
PY - 2023/9/1
Y1 - 2023/9/1
N2 - The human pathogen Pseudomonas aeruginosa has a number of virulence factors at its disposal that play crucial roles in the progression of infection. LasB is one of the major virulence factors and exerts its effects through elastolytic and proteolytic activities aimed at dissolving connective tissue and inactivating host defense proteins. LasB is of great interest for the development of novel pathoblockers to temper the virulence, but access has thus far largely been limited to protein isolated from Pseudomonas cultures. Here, we describe a new protocol for high-level production of native LasB in Escherichia coli. We demonstrate that this facile approach is suitable for the production of mutant, thus far inaccessible LasB variants, and characterize the proteins biochemically and structurally. We expect that easy access to LasB will accelerate the development of inhibitors for this important virulence factor.
AB - The human pathogen Pseudomonas aeruginosa has a number of virulence factors at its disposal that play crucial roles in the progression of infection. LasB is one of the major virulence factors and exerts its effects through elastolytic and proteolytic activities aimed at dissolving connective tissue and inactivating host defense proteins. LasB is of great interest for the development of novel pathoblockers to temper the virulence, but access has thus far largely been limited to protein isolated from Pseudomonas cultures. Here, we describe a new protocol for high-level production of native LasB in Escherichia coli. We demonstrate that this facile approach is suitable for the production of mutant, thus far inaccessible LasB variants, and characterize the proteins biochemically and structurally. We expect that easy access to LasB will accelerate the development of inhibitors for this important virulence factor.
KW - anti-infective agents
KW - LasB
KW - pathoblockers
KW - virulence factors
UR - http://www.scopus.com/inward/record.url?scp=85166184173&partnerID=8YFLogxK
U2 - 10.1002/cbic.202300185
DO - 10.1002/cbic.202300185
M3 - Article
C2 - 37195753
AN - SCOPUS:85166184173
VL - 24
JO - CHEMBIOCHEM
JF - CHEMBIOCHEM
SN - 1439-4227
IS - 17
M1 - e202300185
ER -