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Volume 2

Development of novel anti-bacterials and anti-infectives that target programmed bacterial cell death (BAS anti-microbials)



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EU Contribution

2,016,129 Euro


36 months



Starting date

01 January 2002

cellular assays
drug-resistant bacteria
toxin-antitoxin inhibitors

There is an urgent need for the discovery of novel antibacterial compounds to combat deadly new multi-drug resistant strains. Our targets are the bacterial apoptotic systems (BASs). Since apoptotic systems differ substantially in bacteria and eukaryotes, the BASs and their checkpoint elements constitute an attractive and unexplored family of targets for the development of new antibiotics, which is the essence of our proposal. The novelty of this approach lies in the development of assays to highlight the deregulation of the BASs. Our objective is to engineer bacterial strains for in vivo screening and in vitro biochemical detection methods for the identification of novel compounds that may deregulate the BASs. High-throughput screening (HTS) and detailed structure-activity knowledge of the BASs will be used for synthetic modifications to enhance the specificity of targeting.


The rapid rise in the number of pathogenic bacteria, which have acquired resistance to all commonly used drugs, requires immediate action along new paths. The novelty of our approach is to exploit BASs as a new class of targets for the development of novel anti-microbial compounds and anti-infective interventions. Our aim is to develop cellular assays to monitor the activity of the BASs, and to render them suitable for HTS of chemical libraries and natural compounds collections. Checkpoint elements and the BASs will be assessed as targets in Gram-positive and Gram-negative bacteria. The checkpoint elements and the cellular targets of the toxins will be analysed by employment of genomics and proteomics. Biochemical and biophysical methodologies will be used to characterise the details of toxin-antitoxin protein complexes.


Our project aims to develop cellular assays which monitor DNA-protein and protein-protein interactions found in the BASs in physiologically relevant environments as the first step towards the identification of novel antimicrobial agents. We will engineer bacterial strains for HTS to enable searches for possible antimicrobials that inhibit the formation of macromolecular complexes of the toxin-antitoxin, and toxin-antitoxin with their DNA targets (primary screening). Biochemical assays will be established to ascertain the mode of action of any identified inhibitor and to confirm their function as inhibitors of the BAS. Evaluation and validation of these newly developed assays will allow the search for new inhibitors based on work of this consortium. A natural compound that selectively inhibits the activity of the · toxin-antitoxin will be used to validate our assays. Structural studies on individual toxin-antitoxin components, toxin-antitoxin-DNA and toxin-cellular target complexes will be carried out to provide further information for the industrial partner to exploit a rational approach aimed at finding and improving the desired anti-microbials.


Production of novel in vivo assays to screen for the deregulation of the BASs that are robust, easy to run, cheap, and try to find inhibitors that cross the cell membrane(s). Validation of the highlighted positive hits in vitro. Employment of biochemical and biophysical methodologies yielding data on the details of the interactions of the system components for possible use in rational selection of compounds and tailored drug design. Structural information at medium and high resolution of individual toxin-antitoxin proteins.

Manuel Espinosa
Centro Investigaciones Biológicas
28006 Madrid, Spain
Tel: +34-91-5611 800
Fax: +34-91-5627 518
Juan C. Alonso
Centro Natl. Biotecnología
28049 Madrid, Spain
Tel: +34-91-5854 546
Fax: +34-91-5854 506

Kenn Gerdes
Biochemistry and Molecular Biology
University of Southern Denmark
5230 Odense, Denmark
Tel: +45-6550 2413
Fax: +456550 2467

Dolors Balsa / Andrés G. Fernández
Laboratorios Salvat
08950 Espluges de Llobregat, Spain
Tel: +34-93-3718 600
Fax: +34-93-3732 292

Ehud Gazit
Molecular Microbiology and Biotechnology
Tel-Aviv University
69978 Tel-Aviv, Israel
Tel: +97-2-3640 750
Fax: +97-2-3640 9407

Jerry Wells / Karin Overweg
Institute of Food Research
Norwich Research Park, Colney
NR4 7UA Norwich, United Kingdom
Tel: +44-1603-255 250
Fax: +44-1603-255 037

Wolfram Saenger
Institut für Kristallographie
Freie Universität Berlin
14195 Berlin, Germany
Tel: +49-30-8385 3412
Fax: +49-30-8385 6702

Heinz Welfe
Max-Delbrueck-Center for Molecular Medicine
13092 Berlin-Buch, Germany
Tel: +49-30-9406 2840
Fax: +49-30-9406 2840