X-ray crystal structure of the passenger domain of plasmid encoded toxin(Pet), an autotransporter enterotoxin from enteroaggregative Escherichia coli (EAEC)

Abstract

Autotransporters (ATs) represent a superfamily of proteins produced by a variety of pathogenic bacteria, which include the pathogenic groups of Escherichia coli (E. coli) associated with gastrointestinal and urinary tract infections. We present the first X-ray structure of the passenger domain from the Plasmid-encoded toxin (Pet) a 100 kDa protein at 2.3 Å resolution which is a cause of acute diarrhea in both developing and industrialized countries. Pet is a cytoskeleton-altering toxin that induces loss of actin stress fibers. While Pet (pdb code: 4OM9) shows only a sequence identity of 50 % compared to the closest related protein sequence, extracellular serine protease plasmid (EspP) the structural features of both proteins are conserved. A closer structural look reveals that Pet contains a β-pleaded sheet at the sequence region of residues 181-190, the corresponding structural domain in EspP consists of a coiled loop. Secondary, the Pet passenger domain features a more pronounced beta sheet between residues 135-143 compared to the structure of EspP.

An Acinetobacter trimeric autotransporter adhesin reaped from cells exhibits its nonspecific stickiness via a highly stable 3D structure

“…By contrast, proteins isolated using our enzymatic reaping method are guaranteed to be properly folded and assembled because they are actual parts of the native protein secreted by the original carrier cells. As for classical autotransporters that release their PSDs into the extracellular space, PSDs purified from culture supernatants have been used for biochemical characterization49 or determination of their crystal structures5051. To the best of our knowledge, however, the artificial insertion of a protease recognition site into an autotransporter has never been employed to obtain PSDs for their characterization.…”

Of linkers and autochaperones: an unambiguous nomenclature to identify common and uncommon themes for autotransporter secretion

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Serine Protease Autotransporters of the Enterobacteriaceae (SPATEs): Out and About and Chopping It Up

“…However, only a few SPATEs have been more extensively studied with regards to their biological functions and structural characterization. The crystal structure of the passenger domain of three SPATEs has been determined: EspP from an Escherichia coli O157:H7 strain [6], Hbp (also called Tsh) from an extra-intestinal pathogenic E. coli (ExPEC) strain [7] and Pet from enteroaggregative E. coli (EAEC) strains [8]. Based upon these crystal structures, general models of structure and translocation have been proposed, although, whether such models derived from only a few SPATE structures collectively represent all other SPATEs remains to be determined.…”