ISOLATION, AMINO-ACID-SEQUENCE, AND SYNTHESIS OF DERMASEPTIN, A NOVEL ANTIMICROBIAL PEPTIDE OF AMPHIBIAN SKIN

被引:297
作者
MOR, A
NGUYEN, VH
DELFOUR, A
MIGLIORESAMOUR, D
NICOLAS, P
机构
[1] UNIV PARIS 07,INST JACQUES MONOD,BIOACTIVAT PEPTIDES LAB,2 PL JUSSIEU,F-75251 PARIS 05,FRANCE
[2] INST PASTEUR,MYCOL UNIT,F-75724 PARIS 15,FRANCE
[3] UNIV PARIS 05,PROT LAB,CNRS,URA 1188,F-75270 PARIS 06,FRANCE
关键词
D O I
10.1021/bi00100a014
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A 34-residue antimicrobial peptide named dermaseptin was purified to homogeneity from amphibian skin by a 3-step protocol involving molecular sieve filtration, ion-exchange chromatography, and reversed-phase high-performance liquid chromatography. The complete amino acid sequence of dermaseptin, ALWKTMLKKLGTMALHAGKAALGAAADTISQGTQ, was determined by automated Edman degradation of the peptide and of fragments generated by trypsin. Fast atom bombardment mass spectra of dermaseptin gave a protonated molecular ion m/z 3455.4 which matched the theoretical molecular weight predicted from the amino acid sequence. Dermaseptin was synthesized by the solid-phase method. The synthetic replicate was shown to be indistinguishable from natural dermaseptin with respect to chromatographic properties, amino acid sequence determination, and mass spectrometry analysis. Dermaseptin is a water-soluble, thermostable, and nonhemolytic peptide endowed with highly potent antimicrobial activity against pathogenic fungi at micromolar concentration. Circular dichroism spectra of dermaseptin in hydrophobic media indicated 80% alpha-helical conformation, and predictions of secondary structure suggested that dermaseptin can be configured as an amphiphatic alpha-helix spanning over residues 1-27, a structure that perturbs membrane functions regulating water flux.
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页码:8824 / 8830
页数:7
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