Conjugation of synthetic peptides to proteins: quantitation from S-carboxymethylcysteine released upon acid hydrolysis.
A method described here for conjugating synthetic peptides to carrier proteins provides a convenient method for determining peptide-to-carrier protein ratios. N-Bromoacetyl-containing peptides are reacted in situ with carrier proteins in which the disulfide bonds were reduced with tri-n-butylphosphine. At pH 7-8 and ambient temperature, the newly formed sulfhydryl groups of the carrier protein react exclusively with the bromoacetyl mokiety of the peptide to form conjugates having stable thio ether linkages. Acid hydrolyses of these conjugates release S-carboxymethylcysteine in amounts proportional to the amounts of peptides conjugated and thus allow determination of peptide-to-protein ratios.
Kolodny
N
N
Peptide and Immunochemistry Unit, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892.
Robey
F A
FA
D000085:Acetates; D000818:Animals; D002233:Carbocysteine; D002352:Carrier Proteins; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000031:analogs & derivatives; D006433:Hemocyanins; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008722:Methods; D010455:Peptides / Q000138:chemical synthesis; D011485:Protein Binding; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D013388:Succinimides / Q000138:chemical synthesis
D000085:Acetates; D002352:Carrier Proteins; D010455:Peptides; D012709:Serum Albumin; D013388:Succinimides; D012710:Serum Albumin, Bovine; C049271:N-succinimidyl bromoacetate; C016274:bromoacetate; D002233:Carbocysteine; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D003545:Cysteine