Preparation and utilization of fluorescent synthetic peptides.
In this study, several methods for controlled labelling of synthetic peptides by the use of fluorescent compounds (fluorescein isothiocyanate and dimethylaminonaphthalene sulfonyl chloride) were investigated. The first reagent yielded monofluoresceinated, active compounds only when the peptides lacked lysine residues. Monolabelling of peptides in solution with dimethylaminonaphthalenesulphonyl chloride was hindered by the broad reactivity of the reagent, but was achieved by reacting the fluorochrome on protected resin-bound peptides in solid-phase synthesis. The remarkable stability of the linkage allowed the cleavage of the peptide from the resin and deprotection of side-chain functions without hydrolysis of the labelled group. The binding of antipeptide antibodies to the labelled fragments was then estimated using different techniques.
Chersi
A
A
Regina Elena Institute for Cancer Research, Rome, Italy.
Sezzi
M L
ML
Romano
T F
TF
Evangelista
M
M
Nista
A
A
D000595:Amino Acid Sequence; D003619:Dansyl Compounds; D004797:Enzyme-Linked Immunosorbent Assay; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010455:Peptides; D013050:Spectrometry, Fluorescence; D013861:Thiocyanates
D003619:Dansyl Compounds; D005452:Fluoresceins; D005456:Fluorescent Dyes; D010455:Peptides; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate; C029038:dansyl chloride