Proteomic Techniques for Functional Identification of Bacterial Adhesins

This chapter presents an overview of techniques used in proteomics and describes a functional proteomic approach for identification of a lactoferrin-binding protein of Helicobacter pylori. Proteomics is usually divided into expression proteomics, which includes analysis of protein expression, as well as quantification, functional and structural characterization of proteins in cell lysates and tissues, and cell-map proteomics, which attempts to define all protein-protein interactions and intracellular signaling that occurs in a cell under a given condition. Expression proteomics relies heavily on two-dimensional gel electrophoresis (2D-GE) computer analysis to reveal patterns of protein expression and mass spectrometric analysis of enzymatically cleaved peptides. Cell-mapping proteomics is performed either by affinity purification and identification of protein complexes by mass spectrometry (MS) or by direct DNA readout by yeast two-hybrid, phage display, ribosome display, and RNA-peptide fusion.