Methods and reagents for determining enzyme substrate...

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/00 (2006.01) A61K 38/00 (2006.01) A61K 38/55 (2006.01) C07K 1/04 (2006.01) C12Q 1/37 (2006.01) C12Q 1/42 (2006.01)

Patent

CA 2366671

The invention provides a method for determining a preferred amino acid sequence motif for an active site an associated substrate specificity subsites of an enzyme. In the method of the invention, an enzyme is contacted with an oriented degenerate peptide library, certain peptides within the library which are substrates for the enzyme are bound by the enzyme, and bound peptide/enzyme complexes are separated from unbound peptides. The bound peptides are released from the enzyme and are sequenced. A preferred amino acid sequence motif for the active site and associated specificity subsites is determined based upon the relative abundance of different amino acids residues at each degenerate position. The invention also provides peptides, peptide analogs, and other small molecules which have a variety of uses and can be derived from the present invention.

L'invention concerne un procédé permettant de déterminer un motif préféré de séquence d'acides aminés pour un site actif d'une enzyme et ses sous-sites de spécificité associés. Selon le procédé de l'invention, on met en contact une enzyme avec une banque de peptides dégénérés orientés, certains peptides de la banque étant des substrats pour l'enzyme qui s'y lie, puis on sépare les complexes peptide/enzyme liés et non liés. Les peptides liés sont libérés de l'enzyme puis séquencés. On détermine un motif préféré de séquence d'acides aminés pour le site actif et ses sous-sites de spécificité associés en fonction de l'abondance relative des différents résidus d'acides aminés à chaque position dégénérée. L'invention concerne également des peptides, des analogues peptidiques et d'autres petites molécules convenant pour de nombreuses utilisations et pouvant être dérivés de l'invention.

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