Conversion of alpha-hydroxyalkylated residues in biomolecules using methyltransferases
First Claim
1. A method for targeted conversion of a modified biomolecule, comprisingincubating the modified biomolecule bearing a modifying moiety of formula —
- CH(OH)—
R, wherein R is hydrogen or C1-C12-alkyl, with a cofactor-free directing methyltransferase under conditions compatible with enzymatic activity of the methyltransferase, andobtaining targeted conversion resulting from;
i) covalent removal of said modifying moiety at the target site;
orii) derivatization of said modifying moiety at the target site by covalent coupling of non-cofactor nucleophilic compound(s) of general formula HQ-LX, wherein X represents a functional group or a reporter group attached via a linker L, and Q is selected from S, Se, O, N, or C.
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Abstract
The present invention relates to targeted conversion of alpha-hydroxyalkylated residues in biomolecules in the presence of a directing methyltransferase, namely to targeted removal of the alpha-hydroxyalkyl moieties to give unmodified residues, or targeted derivatization of the alpha-hydroxyalkyl groups by covalent coupling of non-cofactor compounds represented by formula HQ-LX1 wherein X represents a functional group or a reporter group attached via a linker moiety L, and QH is selected from HS—, HSe—, HO—H2N—, HN3 or HCN in the presence of a directing methyltransferase. Further development of the method of targeted conversion comprises methods for targeted labeling a biomolecule and method for detecting hydroxymethylated target sites in a biomolecule according to the present invention.
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Citations
11 Claims
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1. A method for targeted conversion of a modified biomolecule, comprising
incubating the modified biomolecule bearing a modifying moiety of formula — - CH(OH)—
R, wherein R is hydrogen or C1-C12-alkyl, with a cofactor-free directing methyltransferase under conditions compatible with enzymatic activity of the methyltransferase, andobtaining targeted conversion resulting from; i) covalent removal of said modifying moiety at the target site;
orii) derivatization of said modifying moiety at the target site by covalent coupling of non-cofactor nucleophilic compound(s) of general formula HQ-LX, wherein X represents a functional group or a reporter group attached via a linker L, and Q is selected from S, Se, O, N, or C. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
- CH(OH)—
Specification