title: Liquid chromatography-electrospray mass spectrometry of tunicamycin-type antibiotics.
abstract: Several Streptomyces and Clavibacter species produce a family of tunicamycin-like antibiotics (tunicamycins, streptovirudins, corynetoxins, etc.) that inhibit the polyprenol-P:N-acetylhexosamine-1-P translocase family, thus blocking both bacterial cell wall biosynthesis and eukaryotic protein N-glycosylation. The mechanisms of biosynthesis and resistance to these toxins by the producing bacteria are largely unknown. Electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) mass spectrometric techniques are described that structurally assign tunicamycin N-acylated variants in the picomolar range. A voltage gradient across the ESI inlet port was used to generate fragmentation ions that were structurally diagnostic for the tunicamycins. The application of in-line reversed-phase high-performance liquid chromatography-electrospray MS (LC-ESI-MS) resulted in the identification of eight new tunicamycins. Based on these structural assignments a revised nomenclature for tunicamycins is proposed. Application of the LC-ESI-MS methodology to culture supernatants and cellular extracts of the tunicamycin-producing bacterium, Streptomyces lysosuperificus, confirmed tunicamycin production and showed it to be growth-temperature dependent, but did not detect corynetoxins production in culture by phage-infected Clavibacter toxicus.
authors.affiliation: Department of Chemistry, State University of New York, College of Environmental Science and Forestry, 1 Forestry Drive, Syracuse, NY 13210, USA.
authors.forename: B C
authors.initials: BC
authors.lastname: Tsvetanova
authors.forename: N P
authors.initials: NP
authors.lastname: Price
mesh_terms: D000192:Actinomycetales / Q000378:metabolism; D000362:Agar / Q000378:metabolism; D002455:Cell Division; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006017:Glycolipids / Q000378:metabolism; D008956:Models, Chemical; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013302:Streptomyces / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D014415:Tunicamycin / Q000494:pharmacology*
chemical_list: D006017:Glycolipids; C034696:corynetoxins; D014415:Tunicamycin; D000362:Agar