title: Methods for the comprehensive structural elucidation of constitution and stereochemistry of lipopeptides.
abstract: A panel of methods of general suitability for complete structural elucidation of the stereochemistry of cyclopeptides, depsipeptides and lipopeptides is presented and described in detail. The suitability of the proposed methods was exemplified on the lipopeptide poaeamide from Pseudomonas poae. Amino acid configurations have been assigned by direct LC enantiomer separation with Chiralpak ZWIX(+) and were confirmed by GC enantiomer separation on Chirasil L-Val. 3-Hydroxydecanoic acid absolute configuration was analyzed on Chiralpak ZWIX(+) and confirmed by injection on ZWIX(-) which showed opposite elution order. Plenty of d-amino acids have been found in this lipopeptide. It contained in total 5 Leu residues of which one had d-configuration. The position of the d-Leu in the peptide sequence was determined by pepsin and chemical digestions in combination with isolation of diagnostic peptide-fragments and subsequent identification of absolute configurations of the Leu residues. This allowed pinpointing the position of the d-amino acid. The complementarity of the peptide retention profiles on Chiralpak ZWIX column as compared to both RPLC and HILIC suggests its great utility as an alternative peptide separation tool.
authors.affiliation: Institute of Pharmaceutical Sciences, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Gerhardt
authors.affiliation: Institute of Pharmaceutical Sciences, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
authors.forename: Adrian
authors.initials: A
authors.lastname: Sievers-Engler
authors.affiliation: Institute of Pharmaceutical Sciences, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
authors.forename: Ghazaleh
authors.initials: G
authors.lastname: Jahanshah
authors.affiliation: Institute of Pharmaceutical Sciences, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany; Department of Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauß-Allee 11, 93053 Regensburg, Germany.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Pataj
authors.affiliation: Institute of Pharmaceutical Sciences, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany; Department of Pharmaceutical Sciences, University of Perugia, Via del Liceo 1, 06123 Perugia, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Ianni
authors.affiliation: Institute of Pharmaceutical Sciences, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
authors.forename: Harald
authors.initials: H
authors.lastname: Gross
authors.affiliation: Department of Analytical Chemistry, University of Vienna, Waehringer Strasse 38, 1090 Vienna, Austria.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lindner
authors.affiliation: Institute of Pharmaceutical Sciences, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany. Electronic address: michael.laemmerhofer@uni-tuebingen.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Lämmerhofer
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D006868:Hydrolysis; D055666:Lipopeptides / Q000737:chemistry*; D010434:Pepsin A / Q000378:metabolism; D010456:Peptides, Cyclic / Q000737:chemistry; D011487:Protein Conformation; D013237:Stereoisomerism
chemical_list: D000596:Amino Acids; D055666:Lipopeptides; D010456:Peptides, Cyclic; D010434:Pepsin A
keywords: 3-Hydroxyalkanoic acids; Absolute configuration; Amino acids; Chiral separation; Chiralpak ZWIX; Chirasil L-Val; Cyclic lipopeptide; Depsipeptide; Enantioselective chromatography; GC; HPLC