Patent ID: 6207414
Filing Date: 2001-03-27
Classification: C07K,C12N,C12Q,G01N

Abstract:
An in vitro method of sulfating a tyrosine residue on a protein, polypeptide, or peptide, comprising:providing a protein, polypeptide or peptide comprising at least one tyrosine residue;providing a purified tyrosylprotein sulfotransferase;providing 3'-phosphoadenosine 5'-phosphosulfate; andcombining the protein, polypeptide, or peptide with the tyrosylprotein sulfotransferase and the 3'-phosphoadenosine 5'-phosphosulfate under conditions suitable for causing transfer of the sulfate from the 3'-phosphoadenosine 5'-phosphosulfate to the tyrosine of the protein, polypeptide, or peptide thereby forming a sulfated protein, polypeptide or peptide;and wherein the tyrosylprotein sulfotransferase comprises at least one of the sequences selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, amino acids 70-355 of SEQ ID NO:1 or SEQ ID NO:3, amino acids 69-334 of SEQ ID NO:5, amino acids 68-333 of SEQ ID NO:7, amino acids 70-333 of SEQ ID NO:9, amino acids 86-343 of SEQ ID NO:11, and a protein having the activity of a tyrosylprotein sulfotransferase enzyme wherein said protein is encoded by a polynucleotide selected from the group consisting of:(a) a polynucleotide which hybridizes to a nucleic acid sequence comprising nucleotides 82-1194 of SEQ ID NO:2, nucleotides 211-1323 of SEQ ID NO:4, nucleotides 198-1331 of SEQ ID NO:6, nucleotides 157-1287 of SEQ ID NO:8, nucleotides s 66-1208 of SEQ ID NO: 10, or nucleotides 1-1077 of SEQ ID NO:12; and(b) a polynucleotide which hybridizes to a nucleic acid sequence comprising nucleotides 289-1086 of SEQ ID NO:2, nucleotides 418-1215 of SEQ ID NO:4, nucleotides 402-1199 of SEQ ID No:6, nucleotides 358-1155 of SEQ ID NO:8, nucleotides 273-1064 of SEQ ID NO: 10, or nucleotides 256-1029 of SEQ ID NO: 12; andwherein the polynucleotides of (a-b) hybridize under hybridization conditions comprising:incubation at 68.degree. C. overnight followed by washing twice with two times SSC, 0.1% SDS at 22.degree. C., and washing twice with 0.1 times SSC, 0.1% SDS at 50.degree. C.