Engineering a Polyspecific Pyrrolysyl-tRNA Synthetase by a High Throughput FACS Screen.

Adrian Hohl, Ram Karan, Anastassja Gespers (Akal), Dominik Renn, Xuechao Liu, Seema Arun Ghoprade, Michael Groll, Magnus Rueping, Jörg Eppinger

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

The Pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNAPyl are extensively used to add non-canonical amino acids (ncAAs) to the genetic code of bacterial and eukaryotic cells. However, new ncAAs often require a cumbersome de novo engineering process to generate an appropriate PylRS/tRNAPyl pair. We here report a strategy to predict a PylRS variant with novel properties. The designed polyspecific PylRS variant HpRS catalyzes the aminoacylation of 31 structurally diverse ncAAs bearing clickable, fluorinated, fluorescent, and for the first time biotinylated entities. Moreover, we demonstrated a site-specific and copper-free conjugation strategy of a nanobody by the incorporation of biotin. The design of polyspecific PylRS variants offers an attractive alternative to existing screening approaches and provides insights into the complex PylRS-substrate interactions.
Original languageEnglish (US)
JournalScientific reports
Volume9
Issue number1
DOIs
StatePublished - Aug 19 2019

Fingerprint

Dive into the research topics of 'Engineering a Polyspecific Pyrrolysyl-tRNA Synthetase by a High Throughput FACS Screen.'. Together they form a unique fingerprint.

Cite this