Preparation of a verifiable peptide-protein immunogen: direction-controlled conjugation of a synthetic fragment of the monitor peptide with myoglobin and application for sequence analysis.
A useful method for preparing a synthetic peptide-carrying protein for specific antibody production was established. The monitor peptide is a trypsin-sensitive cholecystokinin-releasing peptide purified from rat pancreatic juice on the basis of its stimulatory activity toward pancreatic enzyme secretion. The NH2-terminus fragment of the monitor peptide (residues 1-14) was synthesized by a solid phase method. Cysteine at the COOH terminus of the fragment was conjugated with amino groups of myoglobin using a hetero-bifunctional reagent. Sequence analysis of the fragment-myoglobin conjugate indicated that the peptide/myoglobin conjugation ratio was about 1/1 (mol/mol). Antiserum against the conjugate from a rabbit effectively abolished the stimulatory activity of the monitor peptide in the rat small intestine.
Iwai
K
K
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.
Fukuoka
S
S
Fushiki
T
T
Kido
K
K
Sengoku
Y
Y
Semba
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D000941:Antigens / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D006133:Growth Substances; D007106:Immune Sera / Q000032:analysis; D036341:Intercellular Signaling Peptides and Proteins; D009211:Myoglobin / Q000032:analysis*; D010179:Pancreas / Q000378:metabolism; D010187:Pancreatic Hormones / Q000032:analysis; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000276:immunology; D051381:Rats; D013388:Succinimides; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000032:analysis / Q000276:immunology
D000941:Antigens; D006133:Growth Substances; D007106:Immune Sera; D036341:Intercellular Signaling Peptides and Proteins; D009211:Myoglobin; D010187:Pancreatic Hormones; D010446:Peptide Fragments; C504705:Spink1 protein, rat; D013388:Succinimides; D014359:Trypsin Inhibitor, Kazal Pancreatic; C042424:succinimidyl 4-(4-maleimidophenyl)butyrate