Chemical synthesis of beta-defensins and LEAP-1/hepcidin.
A large and steadily growing subfamily of antimicrobially active peptides of animals and plants is formed by the defensins, which are highly disulfide-bonded, cationic peptides with a molecular mass of about 4 kDa. The synthesis of the human beta-defensins 1 and 2 (hBD-1, hBD-2) as well as of the novel murine beta-defensins 7 and 8 (mBD-7 and mBD-8) is reported. The peptides were synthesized by solid-phase peptide synthesis using fluorenylmethoxycarbonyl chemistry. The linear products were oxidized in the presence of the cysteine/cystine redox system to the biologically active molecules. The correct disulfide connectivity of the resulting cyclic products was partly verified by mass spectrometry and sequence analysis of the fragments obtained after tryptic cleavage. In addition, the recently discovered antimicrobially active human peptide LEAP-1/hepcidin, which contains four disulfide bonds, was successfully synthesized and subsequently oxidized. For Liver-expressed anti microbial peptide (LEAP)-1/hepcidin and hBD-1, the identity of native and synthetic peptides was demonstrated by high-pressure liquid chromatography and capillary electrophoretic analysis. The general synthetic procedure is suitable to rapidly perform the total chemical synthesis of novel fully bioactive defensins, which are expected to be identified soon, as well as of structurally modified analogs.
Klüver
E
E
IPF Pharmaceuticals GmbH, Feodor-Lynen-Strasse 31, D-30625 Hanover, Germany. e.kluever@IPF-Pharmaceuticals.de
Schulz
A
A
Forssmann
W-G
WG
Adermann
K
K
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000737:chemistry; D064451:Hepcidins; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry; D017510:Protein Folding; D011506:Proteins / Q000138:chemical synthesis* / Q000737:chemistry; D023083:beta-Defensins / Q000138:chemical synthesis* / Q000737:chemistry
D023181:Antimicrobial Cationic Peptides; D004220:Disulfides; C578702:HAMP protein, human; C579599:Hamp protein, mouse; D064451:Hepcidins; D009842:Oligopeptides; D011506:Proteins; D023083:beta-Defensins