ATP-dependent leader peptide cleavage by NukT, a bifunctional ABC transporter, during lantibiotic biosynthesis

Mami Nishie, Kouki Shioya, Jun ichi Nagao, Hiroyuki Jikuya, Kenji Sonomoto

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

NukT, a possible ABC transporter maturation and secretion (AMS) protein, may contribute to the cleavage of the leader peptide of NukA, which is the prepeptide of the lantibiotic nukacin ISK-1, and to nukacin ISK-1 transport. In this study, we reconstituted in vitro peptidase activity of the full-length NukT overexpressed in inside-out membrane vesicles of Staphylococcus carnosus TM300. We found that the presence of unusual amino acids in NukA is required for leader peptide cleavage. Furthermore, NukT peptidase activity was inhibited by phenylmethylsulfonyl fluoride, a serine/cysteine protease inhibitor; this finding strongly suggests that NukT, like other AMS proteins, is a cysteine protease. Interestingly, NukT peptidase activity depended on ATP hydrolysis. These results suggest that the N-terminal peptidase domain of NukT may cooperatively function with the C-terminal ATP-binding domain. This is the first in vitro study on lantibiotics that reports the processing mechanism of a full-length bifunctional ABC transporter.

Original languageEnglish
Pages (from-to)460-464
Number of pages5
JournalJournal of Bioscience and Bioengineering
Volume108
Issue number6
DOIs
Publication statusPublished - Dec 1 2009

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Bacteriocins
ATP-Binding Cassette Transporters
Adenosinetriphosphate
Biosynthesis
Protein Sorting Signals
Peptide Hydrolases
Adenosine Triphosphate
Phenylmethylsulfonyl Fluoride
Cysteine Proteinase Inhibitors
Serine Proteinase Inhibitors
Cysteine Proteases
Staphylococcus
Proteins
Hydrolysis
Serine
Amino Acids
Membranes
Amino acids
Processing
nukacin ISK-1

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

ATP-dependent leader peptide cleavage by NukT, a bifunctional ABC transporter, during lantibiotic biosynthesis. / Nishie, Mami; Shioya, Kouki; Nagao, Jun ichi; Jikuya, Hiroyuki; Sonomoto, Kenji.

In: Journal of Bioscience and Bioengineering, Vol. 108, No. 6, 01.12.2009, p. 460-464.

Research output: Contribution to journalArticle

Nishie, Mami ; Shioya, Kouki ; Nagao, Jun ichi ; Jikuya, Hiroyuki ; Sonomoto, Kenji. / ATP-dependent leader peptide cleavage by NukT, a bifunctional ABC transporter, during lantibiotic biosynthesis. In: Journal of Bioscience and Bioengineering. 2009 ; Vol. 108, No. 6. pp. 460-464.
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