GIcNAc phosphotransferase of the lysosomal targeting pathway
AssigneeGenzyme Glycobiology Research Institute, Inc.
InventorWilliam M. Canfield
Life of this patent
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Abstract
The present invention provides nucleotide and amino sequences of the lysosomal targeting pathway enzyme GlcNAc-phosphotransferase, methods of producing and methods of purifying this enzyme.
Claims23
An isolated N-acetylglucosamine-1-phosphotransferase comprising an α-subunit, a β-subunit and a γ-subunit, wherein the α-subunit comprises an amino acid sequence of SEQ ID NO:1 or an amino acid sequence having biological activity of an α-subunit of N-acetylglucosamine-1 -phosphotransferase and which is encoded by an isolated nucleic acid which hybridizes under stringent conditions to the complement of nucleotides 165 to 2948 of SEQ ID NO:4,
- the β-subunit comprises an amino acid sequence of SEQ ID NO:2 or an amino acid sequence having biological activity of a β-subunit of N-acetylglucosamine-1-phosphotransferase and which is encoded by an isolated nucleic acid which hybridizes under stringent conditions to the complement of nucleotides 2949 to 3932 of SEQ ID NO:4, and
- the γ-subunit comprises an amino acid sequence of SEQ ID NO:3 or an amino acid sequence having biological activity of a γ-subunit of N-acetylglucosamine-1-phosphotransferase and which is encoded by an isolated nucleic acid which hybridizes under stringent conditions to the complement of SEQ ID NO:5,
- wherein the stringent conditions comprise washing in 0.2×SSC and 0.1% SDS at 65° C.
An isolated polypeptide, which comprises an amino acid sequence which is encoded by an isolated nucleic acid which hybridizes under stringent conditions to the complement of nucleotides 165 to 2948 of SEQ ID NO:4 and has the activity of a biologically active α-subunit of N-acetylglucosamine-1-phosphotransferase, wherein the stringent conditions comprise washing in 0.2×SSC and 0.1% SDS at 65° C.
An isolated polypeptide, which comprises an amino acid sequence which is encoded by an isolated nucleic acid which hybridizes under stringent conditions to the complement of nucleotides 2949 to 3932 of SEQ ID NO:4 and has the activity of a biologically active β-subunit of N-acetylglucosamine-1 -phosphotransferase, wherein the stringent conditions comprise washing in 0.2×SSC and 0.1% SDS at 65° C.
An isolated polypeptide, which comprises an amino acid sequence which is encoded by an isolated nucleic acid which hybridizes under stringent conditions to the complement of SEQ ID NO:5, and has the activity of a biologically active γ-subunit of N-acetylglucosamine-1-phosphotransferase, wherein the stringent conditions comprise washing in 0.2×SSC and 0.1% SDS at 65° C.
A composition comprising an isolated polypeptide which comprises an amino acid sequence which is encoded by an isolated nucleic acid which hybridizes under stringent conditions to the complement of nucleotides 165 to 2948 of SEQ ID NO:4 and has the activity of a biologically active α-subunit of N-acetylglucosamine-1-phosphotransferase;
- and an isolated polypeptide which comprises an amino acid sequence which is encoded by an isolated nucleic acid which hybridizes under stringent conditions to the complement of nucleotides 2949 to 3932 of SEQ ID NO:4 and has the activity of a biologically active β-subunit of N-acetylglucosamine-1 -phosphotransferase,
- wherein the stringent conditions comprise washing in 0.2×SSC and 0.1% SDS at 65° C.
Dependencies are inferred from the claim text.