Synthesis of proteins by native chemical ligation

Details Synthesis of proteins by native chemical ligation Synthesis of proteins by native chemical ligation

C - Chemistry, Metallurgy

07

K

C07K 1/02 (2006.01) C07K 1/04 (2006.01) C07K 1/08 (2006.01)

Patent

CA 2218620

Proteins of moderate size having native peptide backbones are produced by a method of native chemical ligation. Native chemical ligation employs a chemoselective reaction of two unprotected peptide segments to produce a transient thioester-linked intermediate. The transient thioester-linked intermediate then spontaneously undergoes a rearrangement to provide the full length ligation product having a native peptide bond at the ligation site. Full length ligation products are chemically identical to proteins produced by cell free synthesis. Full length ligation products may be refolded and/or oxidized, as allowed, to form native disulfide-containing protein molecules. The technique of native chemical ligation is employable for chemically synthesizing full length proteins.

L'invention concerne un procédé de ligature chimique endogène permettant de produire des protéines de taille réduite présentant des squelettes peptidiques endogènes. La ligature chimique endogène utilise une réaction chimiosélective de deux segments peptidiques non protégés afin de produire un intermédiaire transitoire lié à un thioester. Ce dernier subit spontanément un réarrangement pour donner un produit de ligature complet comportant une liaison peptidique endogène au niveau du site de ligature. Les produits de ligature complets sont identiques sur le plan chimique aux protéines produites par synthèse sans cellule. Ils peuvent être repliés et/ou oxydés, de façon admise, pour former des molécules de protéines endogènes contenant des bisulfures. La technique de la ligature chimique endogène peut être utilisée pour la synthèse chimique de protéines complètes.

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