A5005617800- Enzyme Production and Characterization
- Biochemical and Structural Characterization
- Oral and gingival health research
- Antibiotic Resistance in Bacteria
- Bacterial biofilms and quorum sensing
- Antimicrobial Peptides and Activities
- Protease and Inhibitor Mechanisms
- Peptidase Inhibition and Analysis
- Oral microbiology and periodontitis research
- Bacillus and Francisella bacterial research
- dental development and anomalies
- Pneumocystis jirovecii pneumonia detection and treatment
- Hemophilia Treatment and Research
- Dupuytren's Contracture and Treatments
- Signaling Pathways in Disease
- Streptococcal Infections and Treatments
- Proteins in Food Systems
- Cancer-related gene regulation
- Biochemical and Molecular Research
- Transgenic Plants and Applications
- Bone and Dental Protein Studies
- Pneumonia and Respiratory Infections
- Epigenetics and DNA Methylation
- Bacteriophages and microbial interactions
- Botulinum Toxin and Related Neurological Disorders
University of Salzburg
2009-2024
Pädagogische Hochschule Salzburg
2023
Clostridial collagenases are among the most efficient enzymes to degrade by far predominant protein in biosphere. Here we present crystal structures of peptidases three clostridial collagenase isoforms (ColG, ColH, and ColT). The comparison unliganded liganded reveals a quaternary subdomain dynamics. In ColH structure, this globular dynamics is modulated an aspartate switch motion that binds catalytic zinc. We further identified calcium binding site proximity Both ions required for full...
In light of the global antimicrobial-resistance crisis, there is an urgent need for novel bacterial targets and antibiotics with modes action. It has been shown that Pseudomonas aeruginosa elastase (LasB) Clostridium histolyticum (Hathewaya histolytica) collagenase (ColH) play a significant role in infection process thereby represent promising antivirulence targets. Here, we report N-aryl-3-mercaptosuccinimide inhibitors target both LasB ColH, displaying potent activities vitro high...
Secreted virulence factors like bacterial collagenases are conceptually attractive targets for fighting microbial infections. However, previous attempts to develop potent compounds against these metalloproteases failed achieve selectivity human matrix metalloproteinases (MMPs). Using a surface plasmon resonance-based screening complemented with enzyme inhibition assays, we discovered an N-aryl mercaptoacetamide-based inhibitor scaffold that showed sub-micromolar affinities toward collagenase...
Antivirulence therapy has become a widely applicable method for fighting infections caused by multidrug-resistant bacteria. Among the many virulence factors produced Gram-negative bacterium Pseudomonas aeruginosa, elastase (LasB) stands out as an important target it plays pivotal role in invasion of host tissue and evasion immune response. In this work, we explored recently reported LasB inhibitor class α-benzyl-N-aryl mercaptoacetamides exploiting crystal structure one compounds. Our...
Infections caused by the Gram-negative pathogen Pseudomonas aeruginosa are emerging worldwide as a major threat to human health. Conventional antibiotic monotherapy suffers from rapid resistance development, underlining urgent need for novel treatment concepts. Here, we report on nontraditional approach combat P. aeruginosa-derived infections targeting its main virulence factor, elastase LasB. We discovered new chemical class of phosphonates with an outstanding in vitro ADMET and PK profile,...
Abstract Clostridial collagenases are used for a broad spectrum of biotechnological applications and represent prime target candidates both therapy diagnosis clostridial infections. In this study, we biochemically characterized the catalytic domains three collagenases, collagenase G (ColG) H (ColH) from Clostridium histolyticum , T (ColT) C. tetani . All protein samples showed activity against synthetic peptidic substrate (furylacryloyl-Leu-Gly-Pro-Ala, FALGPA) with ColH showing highest...
Abstract Collagen is the most abundant protein in higher animals and as such it a valuable source of amino acids carbon for saprophytic bacteria. Due to its unique acid composition triple-helical tertiary structure can however only be cleaved by specialized proteases like collagenases secreted some Among best described bacterial are ColG ColH from Clostridium histolyticum . Many Bacillus species contain homologues clostridial collagenases, which play role infections caused B. cereus Detailed...
Bacterial collagenases are important virulence factors, secreted by several pathogenic Clostridium , Bacillus Spirochaetes and Vibrio species. Yet, the mechanism which these enzymes cleave collagen is not well understood. Based on biochemical mutational studies we reveal that collagenase G (ColG) from Hathewaya histolytica recognizes processes substrates differently depending their nature (fibrillar vs. soluble collagen); distinct dynamic interactions between activator peptidase domain...
Bacterial collagenases differ considerably in their structure and functions. The ColH ColG from Clostridium histolyticum ColA expressed by perfringens are well-characterized that cleave triple-helical collagen, which were therefore termed as ´true´ collagenases. Bacillus cereus (B. cereus) has been added to the collection of true However, molecular characteristics B. less understood. In this study, we identified a secreted collagenase ATCC 14579, is transcriptionally controlled regulon...
Abstract Clostridial collagenases are foe and friend: on the one hand, these enzymes enable host infiltration colonization by pathogenic clostridia, other they valuable biotechnological tools due to their capacity degrade various types of collagen gelatine. However, demand for high-grade preparations exceeds supply origin intricate purification homogeneous isoforms. We present establishment an Escherichia coli expression system a variety constructs collagenase G (ColG) H (ColH) from...
Microbial infections are a significant threat to public health, and resistance is on the rise, so new antibiotics with novel modes of action urgently needed. The extracellular zinc metalloprotease collagenase H (ColH) from Clostridium histolyticum virulence factor that catalyses tissue damage, leading improved host invasion colonisation. Besides major role ColH in pathogenicity, its localisation makes it highly attractive target for development antivirulence agents. Previously, we had found...
In view of the worldwide antimicrobial resistance (AMR) threat, new bacterial targets and anti-infective agents are needed. Since important roles in pathogenesis have been demonstrated for collagenase H G (ColH ColG) from Clostridium histolyticum, Q1 A (ColQ1 ColA) Bacillus cereus represent attractive antivirulence targets. Furthermore, repurposing FDA-approved drugs may assist to tackle AMR crisis was addressed this work. Here, we report on discovery two potent chemically stable inhibitors:...
The alarming increase in global rates of metabolic diseases (MetDs) and their association with cancer risk renders them a considerable burden on our society. interplay environmental genetic factors causing MetDs may be reflected DNA methylation patterns, particularly at non-canonical (non-B) structures, such as G-quadruplexes (G4s) or R-loops. To gain insight into the mechanisms MetD progression, we focused functional analyses intragenic regions two genes, glucokinase (GCK) exon 7...
The catalytic domain of collagenase G from Clostridium histolyticum has been cloned, recombinantly expressed in Escherichia coli and purified using affinity size-exclusion column-chromatographic methods. Crystals the were obtained 0.12 M sodium citrate 23%(v/v) PEG 3350 at 293 K. crystals diffracted to 2.75 Å resolution synchrotron radiation. belong an orthorhombic space group, with unit-cell parameters a = 57, b 109, c 181 Å. This unit cell is consistent presence one molecule per asymmetric...
Despite the progress in surgical techniques and antibiotic prophylaxis, opportunistic wound infections with
Infections caused by the Gram-negative pathogen Pseudomonas aeruginosa are emerging worldwide as a major threat to human health. Conventional antibiotic monotherapy suffers from rapid resistance development, underlining urgent need for novel treatment concepts. Here, we report on non-traditional approach combat P. aeruginosa-derived infections targeting its main virulence factor, elastase LasB. We discovered new chemical class of phosphonates with an outstanding in vitro ADMET and PK...
Antimicrobial resistance is currently one of the serious global public health threats. Unlike conventional antimicrobial drugs, antivirulence agents disarm rather than kill bacterial pathogens and therefore represent an alternative option to skirt problem resistance. Pseudomonas aeruginosa elastase (LasB) Clostridium histolyticum collagenase (ColH) are extracellular proteases which play a critical role in establishment progression respective infection. In this study, we report modulation...
Abstract Collagens form the resilient backbone of extracellular matrix in mammals. Only few proteases are able to digest triple-helical collagen. Clostridial collagenases can efficiently process However, little is known about mechanism bacterial collagenolysis either soluble collagen or multi-hierarchically assembled, insoluble fibers. Here we present a functional analysis distinct roles individual domains collagenase G (ColG) from Hathewaya histolytica. A broad array biochemical,...
Inter-domain movements act as important activity modulators in multi-domain proteins. Here, we present a protocol for inter-domain cross-linking via engineered cysteines. Using collagenase G (ColG) from Hathewaya histolytica model, describe steps the design, expression, purification, and of target protein. We detail system to monitor progress reaction confirm structural integrity purified cross-linked anticipate this be readily adaptable other enzymes. For complete details on use execution...