Unleashing the potential of noncanonical amino acid biosynthesis to create cells with precision tyrosine sulfation
Abstract
Despite the great promise of genetic code expansion technology to modulate structures and functions of proteins, external addition of ncAAs is required in most cases and it often limits the utility of genetic code expansion technology, especially to noncanonical amino acids (ncAAs) with poor membrane internalization. Here, we report the creation of autonomous cells, both prokaryotic and eukaryotic, with the ability to biosynthesize and genetically encode sulfotyrosine (sTyr), an important protein post-translational modification with low membrane permeability. These engineered cells can produce site-specifically sulfated proteins at a higher yield than cells fed exogenously with the highest level of sTyr reported in the literature. We use these autonomous cells to prepare highly potent thrombin inhibitors with site-specific sulfation. By enhancing ncAA incorporation efficiency, this added ability of cells to biosynthesize ncAAs and genetically incorporate them into proteins greatly extends the utility of genetic code expansion methods.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Sep 16, 2022
- Source ID
- 10.1038/s41467-022-33111-4
Entities
People
- Anna Chung
- Han Xiao
- Kuan-lin Wu
- Mengxi Zhang
- Peter G. Wolynes
- Shichao Wang
- Shikai Jin
- Yixian Wang
- Yu Hu
- Yuda Chen
- Zeru Tian
Organizations
- Cancer Prevention and Research Institute of Texas
- Division of Intramural Research, National Institute of Allergy and Infectious Diseases
- National Cancer Institute
- National Institute of General Medical Sciences
- National Science Foundation
- Robert A. Welch Foundation
- United States Department of Defense