An orthogonal seryl-tRNA synthetase/tRNA pair for noncanonical amino acid mutagenesis in Escherichia coli

Claudio Zambaldo; Minseob Koh; Fariborz Nasertorabi; Gye Won Han; Abhishek Chatterjee; Raymond C Stevens; Peter G Schultz

doi:10.1016/j.bmc.2020.115662

An orthogonal seryl-tRNA synthetase/tRNA pair for noncanonical amino acid mutagenesis in Escherichia coli

Bioorg Med Chem. 2020 Oct 15;28(20):115662. doi: 10.1016/j.bmc.2020.115662. Epub 2020 Jul 28.

Authors

Claudio Zambaldo¹, Minseob Koh¹, Fariborz Nasertorabi², Gye Won Han², Abhishek Chatterjee³, Raymond C Stevens⁴, Peter G Schultz⁵

Affiliations

¹ Department of Chemistry and Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037, United States.
² Departments of Biological Sciences and Chemistry, Bridge Institute, Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, United States.
³ Department of Chemistry, Boston College, 2609 Beacon Street, 246B Merkert Chemistry Center, Chestnut Hill, MA 02467, United States. Electronic address: [email protected].
⁴ Departments of Biological Sciences and Chemistry, Bridge Institute, Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, United States. Electronic address: [email protected].
⁵ Department of Chemistry and Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037, United States. Electronic address: [email protected].

Abstract

We report the development of the orthogonal amber-suppressor pair Archaeoglobus fulgidus seryl-tRNA (Af-tRNA^Ser)/Methanosarcina mazei seryl-tRNA synthetase (MmSerRS) in Escherichia coli. Furthermore, the crystal structure of MmSerRS was solved at 1.45 Å resolution, which should enable structure-guided engineering of its active site to genetically encode small, polar noncanonical amino acids (ncAAs).

Keywords: E. coli orthogonality; Genetic code expansion; Non-canonical amino acids; Seryl-tRNA synthetase; X-ray crystallography.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Amino Acids / genetics
Amino Acids / metabolism*
Archaeoglobus fulgidus / enzymology
Escherichia coli / metabolism*
Methanosarcina / enzymology
Protein Engineering
RNA, Transfer / chemistry
RNA, Transfer / metabolism*
Serine-tRNA Ligase / chemistry
Serine-tRNA Ligase / metabolism*

Substances

Amino Acids
RNA, Transfer
Serine-tRNA Ligase

Abstract

Publication types

MeSH terms

Substances

Grants and funding