Biosynthesis of the lantibiotic Pep5. Isolation and characterization of a prepeptide containing dehydroamino acids.
Pep5 is a tricyclic peptide antibiotic which contains the unusual amino acids dehydrobutyrine, lanthionine and 3-methyllanthionine. It is matured from a 60-amino-acid precursor peptide (pre-Pep5) deduced from the sequence of the structural gene pepA. To study the biosynthesis of Pep5 we tried to isolate the primary translation product. We identified a peptide in crude extracts of the Pep5-producing Staphylococcus epidermidis strain using antibodies raised against a synthetic 26-residue peptide representing the leader peptide region of pre-Pep5. The putative precursor was purified by reversed-phase HPLC. The isolated peptide did not react with antibodies directed against a C-terminal fragment of mature Pep5 containing two sulfide bridges. Neither lanthionine nor 3-methyllanthionine was detected in amino acid analysis of the isolated precursor. Its amino acid sequence was identical with the sequence predicted from pepA, but Edman degradation stopped at the first threonine residue of the prolantibiotic region indicating a posttranslational modification at this position. The molecular mass of the isolated peptide was 6575.4 +/- 1.7 Da, determined by ion-spray mass spectrometry. This is in agreement with a molecule being dehydrated at the four threonine and the two serine residues in the propeptide region; such a peptide has a calculated molecular mass of 6576.7 Da. The results strongly suggest that maturation of the lantibiotic Pep5 is initiated by selective dehydration of hydroxyamino acids in the propeptide region of the primary translation product and that thioether ring formation is not closely linked to dehydration.
Weil
H P
HP
Institut für Medizinische Mikrobiologie und Immunologie, Universität Bonn, Federal Republic of Germany.
Beck-Sickinger
A G
AG
Metzger
J
J
Stevanovic
S
S
Jung
G
G
Josten
M
M
Sahl
H G
HG
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000276:immunology; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000276:immunology; D001430:Bacteriocins; D015153:Blotting, Western; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010452:Peptide Biosynthesis; D010455:Peptides / Q000276:immunology; D011498:Protein Precursors / Q000378:metabolism; D021382:Protein Sorting Signals / Q000737:chemistry; D013210:Staphylococcus / Q000235:genetics / Q000378:metabolism*; D014867:Water
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D001430:Bacteriocins; D010455:Peptides; D011498:Protein Precursors; D021382:Protein Sorting Signals; D014867:Water; C054222:lantibiotic Pep5