Specificity of the lipase-specific foldases of gram-negative bacteria and the role of the membrane anchor.
Folding of lipases that are secreted by Pseudomonads and other gram-negative bacteria via the type II secretion pathway is facilitated by dedicated chaperones, called lipase-specific foldases (Lifs). Lifs are membrane-anchored proteins with a large periplasmic domain. The functional interaction between the Lif and its cognate lipase is specific, since the Pseudomonas aeruginosa Lif was found not to substitute for Lifs from Burkholderia glumae or Acinetobacter calcoaceticus. However, the P. aeruginosa Lif was able to activate the lipase from the closely related species P. alcaligenes. Hybrid proteins constructed from parts of the P. aeruginosa and B. glumae Lifs revealed that the C-terminal 138 amino acids of the B. glumae Lif determine the specificity of the interaction with the cognate lipase. Furthermore, the periplasmic domain of the B. glumae Lif was functional when cloned in frame with a cleavable signal sequence, which demonstrates that the membrane anchor is not essential for Lif function in vivo. However, the recombinant Lif was released into the medium, indicating that the function of the membrane anchor is to prevent secretion of the Lif together with the lipase.
Department of Molecular Microbiology, Utrecht University, The Netherlands. m.elkhattabi@bio.uu.nl
M
M
El Khattabi
C
C
Ockhuijsen
W
W
Bitter
K E
KE
Jaeger
J
J
Tommassen
D016954:Acinetobacter calcoaceticus / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D019117:Burkholderia / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics* / Q000378:metabolism*; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids; D017510:Protein Folding; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D001426:Bacterial Proteins; C079999:LipA protein, Bacteria; D008565:Membrane Proteins; D018832:Molecular Chaperones; D008049:Lipase