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Biocomputational Strategies for Microbial Drug Target Identification

The complete genome sequences of about 300 bacteria (mostly pathogenic) have been determined, and many more such projects are currently in progress. The detection of bacterial genes that are non-homologous to human genes and are essential for the survival of the pathogen represent a promis ...

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Methods to Assay Inhibitors of DNA Gyrase and Topoisomerase IV Activities

DNA gyrase and DNA topoisomerase (topo) IV are the bacterial targets of coumarin and quinolone antimicrobial agents. Widespread resistance to clinically important antibiotics such as beta-lactams and macrolides has stimulated the development of novel gyrase and topo IV inhibit ...

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Studies of Enzymes That Cause Resistance to Aminoglycosides Antibiotics

Aminoglycoside antibiotics are highly potent, wide-spectrum bactericidals (1, 2). Bacterial resistance to aminoglycosides, however, is a major problem in the clinical use of aminoglycosides. Enzymatic modification of aminoglycosides is the most frequent resistance mode am ...

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Methods to Identify and Characterize Inhibitors of Bacterial RNA Polymerase

RNA polymerase is essential to the viability of bacteria in all phases of growth and development and is a proven chemotherapeutic target as the cellular target of the rifamycin class of antibiotics. However, despite the characterization of multiple different classes of natural products ...

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A Method to Assay Inhibitors of DNA Polymerase IIIC Activity

The need for new drugs to treat infections caused by antibiotic-resistant bacterial strains has prompted many studies to identify novel targets in pathogenic bacteria. Among the three DNA polymerases expressed by bacteria, one of these, designated pol III, is responsible for DNA replic ...

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Methods to Assay Inhibitors of tRNA Synthetase Activity

Aminoacyl-tRNA synthetases (aa-RS) attracted interest as potential targets for new antibacterial compounds. Most organisms express 20 aa-RSs: one for each amino acid. Aa-RSs are essential proteins in all living organisms. When one aa-RS is inhibited, the corresponding tRNA is not char ...

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Three Methods to Assay Inhibitors of Ribosomal Subunit Assembly

The inhibition of bacterial ribosomal subunit formation is a novel target for translational inhibitors. Inhibition of subunit biogenesis has been shown to be equivalent to the inhibition of protein biosynthesis for many antibiotics. This chapter describes three methods for exami ...

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Inhibition ofChaperone-Dependent Bacterial Ribosome Biogenesis

In Escherichia coli, the molecular chaperone HSP70 (DnaK) is necessary for 30S and 50S ribosomal subunit assembly at temperatures above 37�C. Inhibitors of DnaK should therefore hinder ribosome biogenesis, in addition to all of the other DnaK-dependent cellular functions. An easily te ...

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Assays for the Identification of Inhibitors Targeting Specific Translational Steps

While bacterial protein synthesis is the target of about half of the known antibiotics, the great structural-functional complexity of the translational machinery still offers remarkable opportunities for identifying novel and specific inhibitors of unexploited targets. We ...

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SPARK: A New Peptidyl Transferase Activity Assay

The formation of peptide bonds is the central chemical reaction during protein synthesis and is catalyzed by the peptidyl transferase center residing in the large ribosomal subunit. This active site is composed of universally conserved rRNA nucleosides. The peptidyl transferase ce ...

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The IMGT/HLA Database

The human leukocyte antigen (HLA) complex is located within the 6p21.3 region on the short arm of human chromosome 6 and contains more than 220 genes of diverse function. Many of the genes encode proteins of the immune system and include many highly polymorphic HLA genes. The naming of new HLA genes and all ...

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Searching and Mapping of T-Cell Epitopes, MHC Binders, and TAP Binders

This chapter describes searching and mapping tools of MHCBN database, which is a curated database. It comprises over 23,000 peptide sequences, whose binding affinity with major histocompatibility complex (MHC) or transporter associated with antigen processing (TAP) molecules h ...

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Searching and Mapping of B-Cell Epitopes in Bcipep Database

One of the major challenges in the field of subunit vaccine design is to identify the antigenic regions in an antigen, which can activate B cell. These antigenic regions are called B-cell epitopes. In this chapter, we describe how to use Bcipep, which is a database of experimentally determined linear B ...

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Structural Basis for HLA-A2 Supertypes

The human leukocyte antigen (HLA) alleles are extremely polymorphic among ethnic population, and the peptide-binding specificity varies for different alleles in a combinatorial manner. However, it has been suggested that majority of alleles can be covered within few HLA supertypes, ...

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Searching Haptens, Carrier Proteins, and Anti-Hapten Antibodies

Haptens are small molecules that are usually nonimmunogenic unless coupled to some carrier proteins. The generation of anti-hapten antibodies is important for the development of immunodiagnostics and therapeutics. Recently, our group has developed a database called HaptenDB, ...

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Definition of MHC Supertypes Through Clustering of MHC Peptide-Binding Repertoires

Identification of peptides that can bind to major histocompatibility complex (MHC) molecules is important for anticipation of T-cell epitopes and for the design of epitope-based vaccines. Population coverage of epitope vaccines is, however, compromised by the extreme polymorph ...

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Grouping of Class I HLA Alleles Using Electrostatic Distribution Maps of the Peptide Binding Grooves

Human leukocyte antigen (HLA) molecules involved in immune function by binding to short peptides (8–20 residues) have different sequences in different individuals belonging to distinct ethnic population. Hence, the peptide-binding function of HLA alleles is specific. Class I HLA a ...

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The Classification of HLA Supertypes by GRID/CPCA and Hierarchical Clustering Methods

Biological experiments often produce enormous amount of data, which are usually analyzed by data clustering. Cluster analysis refers to statistical methods that are used to assign data with similar properties into several smaller, more meaningful groups. Two commonly used cluster ...

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Prediction of Peptide-MHC Binding Using Profiles

Prediction of peptide binding to major histocompatibility complex (MHC) molecules is a basis for anticipating T-cell epitopes. Peptides that bind to a given MHC molecule are related by sequence similarity. Therefore, a position-specific scoring matrix (PSSM)—also known as profile ...

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Application of Machine Learning Techniques in Predicting MHC Binders

The machine learning techniques are playing a vital role in the field of immunoinformatics. In the past, a number of methods have been developed for predicting major histocompatibility complex (MHC)-binding peptides using machine learning techniques. These methods allow predict ...

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