C-terminal peptide or protein sequencing reagent and method
AssigneeCity of Hope
InventorsJerome M. Bailey, John E. Shively
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Abstract
A carboxy terminal protein sequencing process is disclosed which utilizes phosphoroisothiocyanatidate for the derivatization step.
Claims14
In a process for the carboxy terminal sequencing of a polypeptide which involves the reaction of said protein or peptide with an isothiocyanate reagent to for a carboxy-terminal thiohydantoin amino acid, the improvement which comprises utilizing as said isothiocyanate reagent, a combination of phosphoroisothiocyanatidate and pyridine.
A process for the carboxy terminal sequencing of a protein or peptide sample which comprises (i) reacting said sample with a reagent consisting essentially of phosphoroisothiocyanatidate and pyridine to form a thiohydantoin derivative of the carboxy terminal amino acid thereof, and
- (ii) reacting said derivatized sample with sodium trimethylsilanolate to release a thiohydantoin amino acid.
In a process for the carboxy terminal sequencing of a polypeptide in which a thiohydantoin derivative of the carboxy terminal amino acid of said polypeptide is formed and said derivative is then cleaved to release a thiohydantoin amino acid, the improvement which comprises reacting said carboxy terminal amino acid with a mixture of phosphoroisothiocyanatidate and pyridine to form said thiohydantoin derivative.
Dependencies are inferred from the claim text.