As with other biomolecules synthesized in biological systems, the stereochemistry and chiral recognition of proteins and peptides are strongly affected by the stereochemistry of their amino acid building blocks. Since the first report on ammonium tartrate enantiomers by Pasteur in 1858 2, the stereochemistry of biomolecules has been a significant topic, as the construction of chiral interfaces and their effect on the behavior of various substances crucially affect various biological and physiological processes 1, 3. The amino acid configuration of the pentapeptide was also determined successfully by subjecting its acid hydrolysates to the Edman reaction followed by HPLC–MS/CD.Ĭhirality, a fundamental attribute of nature, has been found to be ubiquitous it plays a pivotal role in biochemical environments 1. The sequence and configuration of the pentapeptide and up to eight residues of halicylindramide C were successfully analyzed by this method. This method was applied for the detection of a synthetic pentapeptide and a natural depsipeptide (halicylindramide C). These standard PTH derivatives showed well-resolved resolution without interference from byproducts in the ion chromatogram and clear positive/negative CD absorptions when subjected on a reversed phase HPLC–MS system coupled with a CD-2095 HPLC detector. The CD spectra of the derivatives revealed that each pair of the PTH derivatives exhibited the absorption with opposite signs at around 270 nm. Phenylthiohydantoin (PTH) derivatives of 20 pairs of standard d- and l-amino acids were synthesized by the Edman reaction. We report a method for the simultaneous determination of the sequence and absolute configuration of peptide amino acids using a combination of Edman degradation and HPLC–MS/CD.
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