RpeA is an autotransporter protein found in rabbit-specific enteropathogenic Escherichia coli that is involved in colonisation of the rabbit gastrointestinal tract (REPEC). As a member of the proteolytic autotransporter subgroup--serine protease autotransporters of the Enterobacteriaceae (SPATE)--for possessing a characteristic three-domain structure and a serine protease motif in its functional passenger domain, RpeA is a unique existence as 1. it lacks the characteristic auto-catalytic cleavage site between its passenger and β-barrel domain; 2. its passenger domain owns a functionally unclear proline-rich tandem repeat that we hypothesize may be associated with bacterial adherence. We used cellular fractionation and immunofluorescence staining of live RpeA-expressing bacterial cells to show that RpeA is a surface-exposed outer membrane protein. Given its location, further assays will test for potential binding functions in biofilm formation, auto-aggregation and adherence to host cells. We PCR amplified, cloned, and successfully purified as a histidine-tagged recombinant RpeA protein using immobilized metal affinity chromatography with nickel resin combined with size exclusion chromatography. We showed that the purified RpeA passenger domain is a functional protease. Circular dichroism analysis of purified RpeA passenger domain revealed that it is folded and composed of mainly in β-sheet structure. Further functional analysis of RpeA will expand our knowledge of the roles that SPATEs play in their respective pathogens. The passenger domain of RpeA, being proven to be a purifiable folded protein, stands as a significant starting point for crystallization and determination of its tertiary structure.