Genetically encoded tyrosine sulfation of proteins in eukaryotes
AssigneeThe Trustees of Boston College
InventorsAbhishek Chatterjee, James S. Italia
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Abstract
An engineered tyrosyl-tRNA synthetase/tRNA pair that co-translationally incorporates O-sulfotyrosine in response to UAG codons in E. coli and mammalian cells is described herein. This platform enables recombinant expression of eukaryotic proteins homogeneously sulfated at chosen sites.
Claims15
A composition comprising a variant E. coli tyrosyl-tRNA synthetase (EcTyr-RS) wherein the variant EcTyr-RS preferentially aminoacylates an E. coli tyrosyl-tRNA (EctRNAtyr) with a sulfotyrosine analog over the naturally-occurring tyrosine amino acid, wherein the variant EcTyr-RS comprises an amino acid sequence selected from the group consisting of:
- SEQ ID NOS:
- 4-9.
A cell comprising a variant E. coli tyrosyl tRNA synthetase (EcTyr-RS), wherein the variant EcTyr-RS preferentially aminoacylates an E. coli tyrosyl tRNA with a sulfotyrosine analog over the naturally-occurring tyrosine amino acid, and an orthogonal E. coli tyrosyl tRNA (Ec-tRNATyr) as a pair, wherein the variant EcTyr-RS comprises an amino acid sequence selected from the group consisting of:
- SEQ ID NOS:
- 4-9.
A kit for producing a protein or peptide of interest in a cell, wherein the protein or peptide comprises one, or more sulfotyrosine analogs, the kit comprising:
- a. a container containing a polynucleotide sequence encoding an Ec-tRNATyr that recognizes an amber selector codon in a nucleic acid of interest in the cell;
- and
- b. a container containing a variant E. coli tyrosyl tRNA synthetase that preferentially aminoacylates the Ec-tRNATyr with a sulfotyrosyl analog, wherein the EcTry-RS comprises an amino acid sequence selected from the group consisting of:
- SEQ ID NOS:
- 4-9.
Dependencies are inferred from the claim text.