Tyrosylprotein sulfotransferase: purification and molecular cloning of an enzyme that catalyzes tyrosine O-sulfation, a common posttranslational modification of eukaryotic proteins

Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2896-901. doi: 10.1073/pnas.95.6.2896.

Abstract

Tyrosine O-sulfation is a common posttranslational modification of proteins in all multicellular organisms. This reaction is mediated by a Golgi enzyme activity called tyrosylprotein sulfotransferase (TPST) that catalyzes the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to tyrosine residues within acidic motifs of polypeptides. Tyrosine O-sulfation has been shown to be important in protein-protein interactions in several systems. For example, sulfation of tyrosine residues in the leukocyte adhesion molecule P-selectin glycoprotein ligand 1 (PSGL-1) is required for binding to P-selectin on activated endothelium. In this report we describe the purification of TPST from rat liver microsomes based on its affinity for the N-terminal 15 amino acids of PSGL-1. We have isolated human and mouse TPST cDNAs that predict type II transmembrane proteins of 370 amino acid residues with almost identical primary structure. The human cDNA encodes a fully functional N-glycosylated enzyme with an apparent molecular mass of approximately 54 kDa when expressed in mammalian cells. This enzyme defines a new class of Golgi sulfotransferases that may catalyze tyrosine O-sulfation of PSGL-1 and other protein substrates involved in diverse physiologic functions including inflammation and hemostasis.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chromatography, Affinity
  • Cloning, Molecular
  • Eukaryotic Cells / enzymology
  • Humans
  • Male
  • Membrane Glycoproteins / metabolism
  • Mice
  • Microsomes, Liver / enzymology
  • Molecular Sequence Data
  • Phosphoadenosine Phosphosulfate / metabolism
  • Protein Binding
  • Protein Processing, Post-Translational*
  • Rats
  • Rats, Sprague-Dawley
  • Recombinant Proteins / biosynthesis
  • Sequence Homology, Amino Acid
  • Sulfotransferases / biosynthesis
  • Sulfotransferases / genetics*
  • Sulfotransferases / isolation & purification*
  • Tyrosine / metabolism*

Substances

  • Membrane Glycoproteins
  • P-selectin ligand protein
  • Recombinant Proteins
  • Tyrosine
  • Phosphoadenosine Phosphosulfate
  • Sulfotransferases
  • protein-tyrosine sulfotransferase
  • estrone sulfotransferase

Associated data

  • GENBANK/AF038008
  • GENBANK/AF038009