Functional characterization of the trigger factor protein PceT of tetrachloroethene-dechlorinating Desulfitobacterium hafniense Y51.
Desulfitobacterium hafniense strain Y51 dechlorinates tetrachloroethene to cis-1,2-dichloroethene (cis-DCE) via trichloroethene by the action of the PceA reductive dehalogenase encoded by pceA. The pceA gene constitutes a gene cluster with pceB, pceC, and pceT. However, the gene components, except for pceA, still remained to be characterized. In the present study, we characterized the function of PceT. PceT of strain Y51 showed a sequence homology with trigger factor proteins, although it is evolutionally distant from the well-characterized trigger factor protein of Escherichia coli. The PceT protein tagged with 6x histidine was expressed as a soluble form in E. coli. The recombinant PceT fusion protein exhibited peptidyl-proryl cis-trans isomerase activity toward the chromogenic peptide N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide. The PceT fusion protein also exhibited chaperon activity towards the chemically denatured citrate synthase. Immunoprecipitation analysis using antibodies raised against PceA and PceT demonstrated that PceT specifically binds to the precursor form of PceA with an N-terminal twin-arginine translocation (TAT) signal sequence. On the other hand, PceT failed to bind the mature form of PceA that lost the TAT signal sequence. This is the first report in dehalorespiring bacteria, indicating that PceT is responsible for the correct folding of the precursor PceA.
Morita
Yasuyuki
Y
Department of Bioscience and Biotechnology, Kyushu University, Fukuoka, Japan.
Futagami
Taiki
T
Goto
Masatoshi
M
Furukawa
Kensuke
K
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002846:Chromatography, Affinity; D002950:Citrate (si)-Synthase / Q000378:metabolism; D045858:Desulfitobacterium / Q000235:genetics* / Q000378:metabolism*; D004000:Dichloroethylenes / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics; D015870:Gene Expression; D007535:Isomerases / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D005810:Multigene Family; D009842:Oligopeptides / Q000378:metabolism; D019696:Peptidylprolyl Isomerase / Q000235:genetics; D010802:Phylogeny; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013750:Tetrachloroethylene / Q000378:metabolism*; D014241:Trichloroethylene / Q000378:metabolism
D001426:Bacterial Proteins; D004000:Dichloroethylenes; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D014241:Trichloroethylene; C043307:succinyl-alanyl-alanyl-prolyl-phenylalanine-4-nitroanilide; D002950:Citrate (si)-Synthase; D007535:Isomerases; C403324:trigger factor, E coli; D019696:Peptidylprolyl Isomerase; D013750:Tetrachloroethylene; C035384:1,2-dichloroethylene