Biochemical structural analysis of the lantibiotic mutacin II

Robert E. Krull, Ping Chen, Jan Novak, Marion Kirk, Stephen Barnes, John Baker, N. Rama Krishna, Page W. Caufield

Research output: Contribution to journalArticle

Abstract

Mutacin II is a post-translationally modified lantibiotic peptide secreted by Streptococcus mutans T8, which inhibits the energy metabolism of sensitive cells. The deduced amino acid sequence of promutacin II is NRWWQGVVPTVSYECRMNSWQHVFTCC, which is capable of forming three thioether bridges. It was not obvious, however, how the three thioether bridges are organized. To examine the bridging, the cyanogen bromide cleavage products of mutacin II and its variants generated by protein engineering, C15A, C26A, and C15A/C26A, were analyzed by mass spectrometry. Analysis of the wild type molecule and the C15A variant excluded several possibilities and also indicated a high fidelity of formation of the thioether bridges. This allowed us to further resolve the structure by analysis (mass spectrometry and tandem mass spectrometry) of the cyanogen bromide cleavage fragments of the C26A and C15A/C26A mutants. Nuclear magnetic resonance analysis established the presence of one and two dehydrobutyrine residues in mutacin II and the C15A variant, respectively, thus yielding the final structure. The results of this investigation showed that the C-terminal part contains three thioether bridges connecting Cys residues 15, 26, and 27 to Ser/Thr residues 10, 12 and 19, respectively, with Thr25 being modified to dehydrobutyrine.

Original languageEnglish (US)
Pages (from-to)15845-15850
Number of pages6
JournalJournal of Biological Chemistry
Volume275
Issue number21
DOIs
StatePublished - May 26 2000

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Bacteriocins
Sulfides
Structural analysis
Mass spectrometry
Cyanogen Bromide
Tandem Mass Spectrometry
Protein Engineering
Streptococcus mutans
Energy Metabolism
Amino Acid Sequence
Mass Spectrometry
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Amino Acids
Peptides
Molecules
mutacin II
Proteins
dehydrobutyrine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Krull, R. E., Chen, P., Novak, J., Kirk, M., Barnes, S., Baker, J., ... Caufield, P. W. (2000). Biochemical structural analysis of the lantibiotic mutacin II. Journal of Biological Chemistry, 275(21), 15845-15850. https://doi.org/10.1074/jbc.275.21.15845

Biochemical structural analysis of the lantibiotic mutacin II. / Krull, Robert E.; Chen, Ping; Novak, Jan; Kirk, Marion; Barnes, Stephen; Baker, John; Krishna, N. Rama; Caufield, Page W.

In: Journal of Biological Chemistry, Vol. 275, No. 21, 26.05.2000, p. 15845-15850.

Research output: Contribution to journalArticle

Krull, RE, Chen, P, Novak, J, Kirk, M, Barnes, S, Baker, J, Krishna, NR & Caufield, PW 2000, 'Biochemical structural analysis of the lantibiotic mutacin II', Journal of Biological Chemistry, vol. 275, no. 21, pp. 15845-15850. https://doi.org/10.1074/jbc.275.21.15845
Krull RE, Chen P, Novak J, Kirk M, Barnes S, Baker J et al. Biochemical structural analysis of the lantibiotic mutacin II. Journal of Biological Chemistry. 2000 May 26;275(21):15845-15850. https://doi.org/10.1074/jbc.275.21.15845
Krull, Robert E. ; Chen, Ping ; Novak, Jan ; Kirk, Marion ; Barnes, Stephen ; Baker, John ; Krishna, N. Rama ; Caufield, Page W. / Biochemical structural analysis of the lantibiotic mutacin II. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 21. pp. 15845-15850.
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