Extraction and Electrophoretic Analysis of Bacterial Lipopolysaccharides and Outer Membrane Proteins

Abstract

Lipopolysaccharides (LPS) (or lipooligosaccharides [LOS], which lack the O-antigen side chains characteristic of LPS), and outer membrane proteins (OMP) are major cell-surface molecules in the outer membrane (OM) of gram-negative bacteria. The LPS is responsible for causing endotoxic shock in infected hosts and, in conjunction with some OMPs, provides protection to the bacterium against host innate immune defenses and attachment to host cells. Electrophoretic analysis can provide valuable information regarding the size, number, and variability of LPS/LOS and OMP components between bacterial strains and mutants, which aids in understanding the basic biology and virulence factors of a particular species. Furthermore, highly purified extracts are normally not required if only electrophoretic analysis is to be done, and various methods have been established for such procedures. Here, we review ameliorated procedures for fast and convenient extraction of LPS/LOS and protein-enriched outer membranes (PEOM) for optimal electrophoretic resolution. Specifically, we will describe the phenol-based micro-method for LPS/LOS extraction, a differential extraction procedure with sodium lauryl sarcosinate for PEOM, and gel preparation for electrophoretic analysis of LPS/LOS samples in detail. Graphic abstract: Workflow for the preparation and analysis of LPS/LOS and PEOM.

Keywords: Lipooligosaccharide (LOS); Lipopolysaccharide (LPS); Outer membrane protein (OMP); Phenol-water microextraction; Protein-enriched outer membranes (PEOM); Silver staining; Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); Sodium lauryl sarcosinate.