Studying Protein Tyrosine O-Sulfation in Mammalian Cells with Genetically Encoded Sulfotyrosine

Xinyuan He, Jiantao Guo, Wei Niu

Research output: Contribution to journalArticlepeer-review

Abstract

Protein tyrosine O-sulfation (PTS) is a post-translational modification (PTM) that occurs exclusively on secreted and membrane-bound proteins. It participates in a wide range of important biological processes and is involved in the development of many diseases. The biomedical importance of PTS can only be fully unveiled when the right chemical/biological tools are available. This article outlines the steps for using an engineered Escherichia coli tyrosyl-tRNA synthetase to genetically encode sulfotyrosine (sTyr) for biological investigations of PTS in mammalian cells. Two basic protocols are described to demonstrate this methodology, including the site-specific incorporation of sTyr into eGFP protein in HEK293T cells and the functional study of an sTyr-containing CXCR4 protein using a calcium mobilization assay.

Original languageEnglish (US)
Article numbere301
JournalCurrent Protocols
Volume1
Issue number11
DOIs
StatePublished - Nov 2021

Keywords

  • CXCR4
  • calcium mobilization
  • genetic code expansion
  • protein tyrosine O-sulfation (PTS)
  • sulfotyrosine incorporation

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Health Informatics
  • Medical Laboratory Technology

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