Design, synthesis, tandem mass spectrometric sequencing and biological activity of NGF mimetics.
Nine low molecular weight nerve growth factor (NGF)-like peptides have been designed to mimic the putative receptor-binding epitope of NGF defined by two beta-hairpin loops. Eight different spacers were used as variable links between the beta-loop amino acid residues, which from mutagenesis experiments were found to play an important role in the biological activity of NGF. These spacers were amino acids, natural or non-natural, differing in length (5-13 A) and polarity. The peptides were synthesized via the Fmoc solid-phase peptide synthesis and purified by reversed-phase HPLC. Their primary sequences were analyzed by a combination of automated Edman degradation and mass spectrometry. The peptides were tested using two different biological assays, the fibre outgrowth from chick embryonic sympathetic ganglia and the PC12 cell differentiation assay. Weak antagonistic effects could be observed for some peptides.
Estenne-Bouhtou
G
G
Department of Organic Pharmaceutical Chemistry, Uppsala Biomedical Centre, Uppsala University, Sweden.
Kullander
K
K
Karlsson
M
M
Ebendal
T
T
Hacksell
U
U
Luthman
K
K
D000313:Adrenal Medulla / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002642:Chick Embryo; D002851:Chromatography, High Pressure Liquid; D018360:Crystallography, X-Ray; D005728:Ganglia, Sympathetic / Q000502:physiology*; D008853:Microscopy; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000737:chemistry* / Q000378:metabolism; D016501:Neurites / Q000187:drug effects / Q000502:physiology; D016716:PC12 Cells; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D012995:Solubility; D018629:Spectrometry, Mass, Secondary Ion
D009414:Nerve Growth Factors; D010455:Peptides