Synthesis and characterization of the human CC chemokine HCC-2.
Human CC chemokine 2 (HCC-2) is a novel member of the chemokine peptide family that induces chemotaxis of monocytes, T lymphocytes and eosinophils via activation of the CCR-1 and CCR-3 receptors. Fmoc chemistry was optimized and used to synthesize the biologically active 66-residue peptide HCC-2-(48-113). Introduction of the three disulfide bonds was achieved by oxidative folding in the presence of the redox system cysteine/cystine. Alternatively, a semiselective approach utilizing a mixed Acm/Trt protection scheme for disulfide formation was applied. It was found that, without participation of the two HCC-2-specific cysteine residues in positions 64 and 104, the two typical chemokine disulfides are formed predominantly during oxidative folding. In addition, the mutant [Ala64,104]HCC-2-(48-113) lacking the third disulfide bond that discriminates HCC-2 from most other chemokines was synthesized. For disulfide bond formation, oxidative folding was compared with the use of Acm/Trt protection. HCC-2-(48-113) and the mutant [Ala64,104]HCC-2-(48-113) were further analyzed by CD and one-dimensional 1H NMR-spectroscopy. Both peptides adopt a similar stable secondary and tertiary structure in solution.
Escher
S E
SE
Niedersächsisches Institut für Peptid-Forschung, Hannover, Germany.
Sticht
H
H
Forssmann
W G
WG
Rösch
P
P
Adermann
K
K
D000595:Amino Acid Sequence; D019742:Chemokines, CC / Q000737:chemistry*; D002942:Circular Dichroism; D006801:Humans; D019402:Macrophage Inflammatory Proteins; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D015846:Monokines; D010084:Oxidation-Reduction; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D013056:Spectrophotometry, Ultraviolet
C108814:CCL15 protein, human; D019742:Chemokines, CC; D019402:Macrophage Inflammatory Proteins; D015846:Monokines