6533b86dfe1ef96bd12c965f
RESEARCH PRODUCT
High-performance liquid chromatographic separation of modified and native melittin following transglutaminase-mediated derivatization with a dansyl fluorescent probe.
Jean DufourcqConcepción AbadLorenzo BracoEnrique Pérez-payásubject
Resolution (mass spectrometry)Tissue transglutaminaseGuinea PigsMolecular Sequence DataPeptideBiochemistryMelittinAnalytical Chemistrychemistry.chemical_compoundCadaverineAmphiphileAnimalsAmino Acid SequenceDerivatizationChromatography High Pressure Liquidchemistry.chemical_classificationDansyl CompoundsChromatographyTransglutaminasesbiologyChemistryElutionOrganic ChemistryGeneral MedicineFluorescenceMelittenSpectrometry Fluorescencebiology.proteinChromatography GelSpectrophotometry Ultravioletdescription
Abstract The 26-amino acid linear, amphiphilic peptide melittin was enzymatically modified with the fluorescent probe monodansylcadaverine using guinea pig liver transglutaminase and a fluorescent derivative of stoichiometry 1:1 was obtained. Reversed-phase and size-exclusion high-performance liquid chromatographic modes were tested in order to resolve the labelled peptide and native species. The influence of several operational variables was analysed and the elution conditions were optimized so that a satisfactory resolution could be achieved in both instances in a rapid, easy manner. Both chromatographic modes offer the possibility of accurate monitoring of the time course of the enzyme-mediated conversion and, more interestingly, can be applied to the semi-preparative purification of the labelled peptide.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1991-07-01 | Journal of chromatography |