Acetic anhydride activation for c-terminal protein sequencing

Details Acetic anhydride activation for c-terminal protein sequencing Acetic anhydride activation for c-terminal protein sequencing

G - Physics

01

N

G01N 33/68 (2006.01) C07K 1/12 (2006.01)

Patent

CA 2197304

A method is provided for C-terminal sequencing of a protein or peptide. An important feature of the method is the formation of an oxazolone moiety at the C-terminus of a protein or peptide by treatment with acetic anhydride under basic conditions followed by conversion of the oxazolone to a thiohydantoin moiety by treatment with thiocyanate under acidic conditions. Yields of thiohydantoin are further enhanced by delivering thiocyanate as the conjugate acid of a sterically hindered alkylammonium cation.

Procédé de séquençage C-terminal d'une protéine ou d'un peptide dont l'un des traits importants est la formation au niveau de la terminaison C de la protéine ou du peptide, d'une fraction oxazolone résultant d'un traitement à l'anhydride acétique dans des conditions basiques, suivi par une conversion de l'oxazolone en une fraction thiohydantoïne par traitement par un thiocyanate dans des conditions acides. Les rendements en thiohydantoïne sont encore améliorés lorsque le thiocyanate est produit comme acide conjugué d'un cation d'alkylamonium à encombrement stérique.

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