Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or...
Reexamination Certificate
1998-02-19
2001-06-12
Borin, Michael (Department: 1642)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
C435S194000
Reexamination Certificate
active
06245502
ABSTRACT:
This application claims benefit of European Patent Application No. 97102616.6, filed Feb. 19, 1997, incorporated herein by reference it its entirety.
1. INTRODUCTION
The invention relates to a drug target system for the identification of novel and highly specific antimicrobials. In particular, the present invention relates to screening assays to identify antimicrobial agents which uncouple or inhibit the bacterial phosphotransferase system.
2. BACKGROUND OF THE INVENTION
Infectious diseases affect all age groups and cause one- third of all death worldwide. Respiratory infections, diarrheal diseases and tuberculosis, neonatal tetanus and whooping cough are the most prominent among the bacterial infections. Infectious disease are still rising due to population growth, urbanization, and not least due to the widespread use of antibiotics that led to the development of antibiotic resistance. The increasing incidence of bacterial drug-resistance requires that new targets of antimicrobial therapy be identified and that inhibitors of these systems be discovered.
3. SUMMARY OF THE INVENTION
The present invention relates to a drug target system for novel and specific antimicrobials which modulate the bacterial phosphotransferase system. In particular, the present invention relates to the identification of agents which uncouple or inhibit the bacterial phosphotransferase system. In a preferred embodiment of the present invention, the assays to identify antimicrobial agents which
a) use a phosphoryl group acceptor which is not part of the phosphotransferase system and preferably hydrolyse spontaneously or under the influence of non- specific endogenous phosphohydrolases thereby uncoupling the phosphotransferase system, or
b) inhibit the phosphotransferase system activity by blocking or inactivating at least one of the enzymes of the phosphotransferase system.
The present invention relates to screening assays to identify potential antimicrobial agents which screen for those agents which utilize a phosphoryl group that is not part of the bacterial phosphotransferase system, or agents which hydrolyse, spontaneously or under the influence of non- specific endogenous phosphohydrolases, thereby uncoupling the system. The present invention further relates to screening assays to identify agents which inhibit at least one of the enzymes of the phosphotransferase system.
Furthermore, the invention relates to novel antimicrobials identified by using said drug target system.
REFERENCES:
patent: 5698428 (1997-12-01), Abo
patent: 5700675 (1997-12-01), Rubin
Ammer et al., 1979, “Phosphorylation of streptozotocin during uptake via the phosphoenolpyruvate: sugar phosphotransferase system inEscherichia coli”, Antimicrob. Agents Chemother. 16:801-807.
Amster-Choder and Wright, 1993, “Transcriptional regulation of the bgl operon ofEscherichia-coliinvolves phosphotransferase system-mediated phosphorylation of a transcriptional antiterminator”, J. Cell. Bioch. 51:83-90.
Buhr et al., 1992 “The glucose transporter ofEscherichia coli.Mutants with impaired translocation activity that retain phosphorylation activity”, J. Biol. Chem. 267:3847-3851.
Christian et al., 1992, “Simplified methods for construction, assessment and rapid screening of peptide libraries in bacteriophages ”, J. Mol. Biol. 227:711-718.
Curtiss III and Kelly, 1987, “Salmonella typhimuriumdeletion mutants lacking adenylate cyclase and cyclic AMP receptor protein are avirulent and immunogenic”, Infect. Immun. 55:30355-3043.
Curtiss III et al., 1988, “AvirulentSalmonella typhimurium&Dgr;cya &Dgr;crp oral vaccine strains expressing a streptococcal colonization and virulence antigen”, Vaccine 6:155-160.
Eberstadt et al., 1966, “Solution structure of the IIB domain of the glucose transporter ofEscherichia coli”, Biochemistry 35:11286-11292.
Erni et al., 1982, “Bacterial phosphotransferase system. Solubilization and purification of the glucose-specific enzyme II from membranes ofSalmonella typhimurium”, J. Biol. Chem. 257:13726-13730.
Erni, 1992, “Group translocation of glucose and other carbohydrates by the bacterial phosphotransferase system”, Int. Rev. Cytol. 137A:127-148.
Hurley et al., 1993, “Structure of the regulatory complex ofEschierichia coliIIIGlcwith glycerol kinase”, Science 259:673-677.
Jia et al., 1993, “Active-centre torsion-angle strain revealed in 1.6 angstrom-resolution structure of histidine-containing phosphocarrier protein”, Nature 361:94-97.
Jia et al., 1993, “The 2.0-angstrom resolution structure ofEscherichia colihistidine-containing phosphocarrier protein HPr -A redetermination”, J. Biol. Chem. 268:22490-22501.
Kalbitzer and Hengstenberg, 1993, “The solution structure of the histidine-containing protein (Hpr) fromStaphylococcus-aureusas determined by 2-dimensional H-1-NMR spectroscopy”, Eur. J. Biochem. 216:205-214.
Kelly et al., 1992, “Characterization and protective properties of attenuated mutants ofSalmonella choleraesuis”Infect. Immun. 60:4881-4890.
Lengeler et al., 1994, “Enzymes II of the phosphoenolpyruvate-dependent phosphotransferase systems: their structure and function in carbohydrate transport”, Biochem. Biophys. Acta 1188:1-28.
Liao et al., 1996, “The first step in sugar transport: crystal structure of the amino terminal domain of enzyme I of theE. coliPEP: sugar phosphotransferase system and a model of the phosphotransfer complex with HPr”, Structure 4:861-872.
Lux et al., 1995, “Coupling the phosphotransferase system and the methyl-accepting chemotaxis protein-dependent chemotaxis signaling pathways ofEscherichia coli”, Proc. Natl. Acad. Sci. USA 92:11583-11587.
Licalsi et al., 1991, “Sugar transport by the bacterial phosphotransferase system. Structural and thermodynamic domains of Enzyme I of Salmonella typhimurium”, J. Biol. Chem. 266:19519-19527.
Mao et al., 1995, “Functional reconstitution of the purified mannose phosphotransferase system ofEscherichia coli.into phospholipid vesicles”, J. Biol. Chem. 270:5258-5265.
Martin,-Verstraete et al., 1994 “Interactions of wild-type and truncated LevR ofBacillus subtiliswith the upstream activating sequence of the levanase operon”, J. Biol. 241:178-192.
Mayahara et al., 1967, “The new lead citrate method for the ultrachemical demonstration of activity of non-specific alkaline phosphatase”, Histochemie 11:88.
Meadow et al.,1990, “The bacterial phosphoenolpyruvate:glycose phosphotransferase system”, Annu. Rev. Biochem. 59:497-542.
Meins et al., 1993, Cysteine phosphorylation of the glucose transporter ofEscherichia coli.J. Biol. Chem. 268, 11604-11609.
Mukhija and Erni, 1996, “Purification by Ni2 +affinity chromatography, and functional reconstitution of the transporter for N-acetylglucosamine ofEscherichia coli”, J. Biol. Chem. 271:14819-14824.
Nishikawa et al., 1995, “Catabolite repression of the adhesion of Vero cytotoxin-producing Escherichia coli sero-groups 01557 an 0111”, Microb. Pathog. 18:223-229.
Postma et al., 1993, “Phosphoenolpyruvate: carbohydrate phosphotransferase systems in bacteria”, Microbiol. Rev. 57:543-594.
Powell et al., 1995, “Novel proteins of the phosphotransferase system encoded within therpoNoperon ofEschericia coli”, J. Biol. Chem. 270:4822-4839.
Rajagopal et al., 1994, “Structural consequences of histidine phosphorylation: NMR characterization of the phosphohistidine form of histidine-containing protein fromBacillus subtilisandEscherichia coli”, Biochemistry 33:15271-15282.
Reizer et al., 1993, “Sequence analyses and evolutionary relationships among the energy-coupling proteins enzyme-I and HPr of the bacterial phosphoenolpyruvate-sugar phosphotransferase system”, Protein Sci. 2:506-521.
Roossien et al., 1983, “A simple procedure for the synthesis of [32P ]phosphosphoenolpyruvate via the pyruvate kinase exchange reaction at equilibrium”, Biochim. Biophsy. Acta. 760:185-187.
Saier, 1993, “Regulatory interactions involving the proteins of the phosphotransferase system in enteric bacteria”, J. Cell. Biochem. 51:62-68.
Saier and Reizer, 1994, “The bacterial phosphotransferase system: New frontiers 30 years later”, Molecular
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