Microbe sample from Escherichia coli

BioSample: SAMN11037806; Sample name: C41 delta ompF delta acrB; SRA: SRS442251

Improving the purification of membrane proteins from Escherichia coli C41(DE3)

The overexpression and purification of membrane proteins to high purity and homogeneity is a challenging task. Over time, several strains have been developed that decrease the toxic side-effects and thus result in higher bio mass and protein yield. However, two major contaminants have been identified in membrane protein preparations from E. coli: the outer membrane porin OmpF and AcrB, which is part of a tripartite efflux pump. Both proteins crystallise from low concentrations and diverse conditions, which make them a major problem, especially in membrane protein crystallography. In this study, we present a C41(DE3)-derived expression strains that is depleted of these two proteins

Identification of Protein Secretion Systems in Bacterial Genomes Using MacSyFinder.

International audienceProtein secretion systems are complex molecular machineries that translocate proteins through the outer membrane, and sometimes through multiple other barriers. They have evolved by co-option of components from other envelope-associated cellular machineries, making them sometimes difficult to identify and discriminate. Here, we describe how to identify protein secretion systems in bacterial genomes using MacSyFinder. This flexible computational tool uses the knowledge stemming from experimental studies to identify homologous systems in genome data. It can be used with a set of predefined models-"TXSScan"-to identify all major secretion systems of diderm bacteria (i.e., with inner and with LPS-containing outer membranes). For this, it identifies and clusters colocalized components of secretion systems using sequence similarity searches with hidden Markov model protein profiles. Finally, it checks whether the genetic content and organization of clusters satisfy the constraints of the model. TXSScan models can be customized to search for variants of known systems. The models can also be built from scratch to identify novel systems. In this chapter, we describe a complete pipeline of analysis, including the identification of a reference set of experimentally studied systems, the identification of components and the construction of their protein profiles, the definition of the models, their optimization, and, finally, their use as tools to search genomic data

Secretion systems in Gram-negative bacteria: structural and mechanistic insights

Bacteria have evolved a remarkable array of sophisticated nanomachines to export various virulence factors across the bacterial cell envelope. In recent years, considerable progress has been made towards elucidating the structural and molecular mechanisms of the six secretion systems (types I–VI) of Gram-negative bacteria, the unique mycobacterial type VII secretion system, the chaperone–usher pathway and the curli secretion machinery. These advances have greatly enhanced our understanding of the complex mechanisms that these macromolecular structures use to deliver proteins and DNA into the extracellular environment or into target cells. In this Review, we explore the structural and mechanistic relationships between these single- and double-membrane-embedded systems, and we briefly discuss how this knowledge can be exploited for the development of new antimicrobial strategies