biostudies-literatureAnlauf MTDeutsche ForschungsgemeinschaftHeinrich-Heine-Universität Düsseldorf8994https://www.ebi.ac.uk/biostudies/studies/S-EPMC11026475biostudies-literatureUnknown14(1)Type I secretion systems (T1SS) facilitate the secretion of substrates in one step across both membranes of Gram-negative bacteria. A prime example is the hemolysin T1SS which secretes the toxin HlyA. Secretion is energized by the ABC transporter HlyB, which forms a complex together with the membrane fusion protein HlyD and the outer membrane protein TolC. HlyB features three domains: an N-terminal C39 peptidase-like domain (CLD), a transmembrane domain (TMD) and a C-terminal nucleotide binding domain (NBD). Here, we created chimeric transporters by swapping one or more domains of HlyB with the respective domain(s) of RtxB, a HlyB homolog from Kingella kingae. We tested all chimeric transporters for their ability to secrete pro-HlyA when co-expressed with HlyD. The CLD proved to be most crScientific reportsType 1 secretion necessitates a tight interplay between all domains of the ABC transporter.PMC11026475INST 208/761-1 FUGGCRC 1208 project A01Grant number 417919780Reiners JSpitz OBilsing FLSmits SHJSchmitt LAnlauf MTHachani EfalseType 1 secretion necessitates a tight interplay between all domains of the ABC transporter.Type I secretion systems (T1SS) facilitate the secretion of substrates in one step across both membranes of Gram-negative bacteria. A prime example is the hemolysin T1SS which secretes the toxin HlyA. Secretion is energized by the ABC transporter HlyB, which forms a complex together with the membrane fusion protein HlyD and the outer membrane protein TolC. HlyB features three domains: an N-terminal C39 peptidase-like domain (CLD), a transmembrane domain (TMD) and a C-terminal nucleotide binding domain (NBD). Here, we created chimeric transporters by swapping one or more domains of HlyB with the respective domain(s) of RtxB, a HlyB homolog from Kingella kingae. We tested all chimeric transporters for their ability to secrete pro-HlyA when co-expressed with HlyD. The CLD proved to be most cr2024-01-01T00:00:00Z2024 Apr2026-06-01T09:18:59.485Z2026-04-08T11:02:33.6ZS-EPMC110264753863767810.1038/s41598-024-59759-0