Structural Basis of Furan–Amino Acid Recognition by a Polyspecific Aminoacyl-tRNA-Synthetase and its Genetic Encoding in Human Cells

Schmidt, Moritz J.; Weber, Annemarie; Pott, Moritz; Welte, Wolfram; Summerer, Daniel

Structural Basis of Furan–Amino Acid Recognition by a Polyspecific Aminoacyl-tRNA-Synthetase and its Genetic Encoding in Human Cells

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Datum

2014

Publikationstyp

Zeitschriftenartikel

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ChemBioChem. 2014, 15(12), pp. 1755-1760. ISSN 1439-4227. eISSN 1439-7633. Available under: doi: 10.1002/cbic.201402006

Zusammenfassung

The site-selective introduction of photo-crosslinking groups into proteins enables the discovery and mapping of weak and/or transient protein interactions with high spatiotemporal resolution, both in vitro and in vivo. We report the genetic encoding of a furan-based, photo-crosslinking amino acid in human cells; it can be activated with red light, thus offering high penetration depths in biological samples. This is achieved by activation of the amino acid and charging to its cognate tRNA by a pyrrolysyl-tRNA-synthetase (PylRS) mutant with broad polyspecificity. To gain insights into the recognition of this amino acid and to provide a rationale for its polyspecificity, we solved three crystal structures of the PylRS mutant: in its apo-form, in complex with adenosine 5′-(β,γ-imido)triphosphate (AMP-PNP) and in complex with the AMP ester of the furan amino acid. These structures provide clues for the observed polyspecificity and represent a promising starting point for the engineering of PylRS mutants with further increased substrate scope.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Schlagwörter

amino acids, crosslinking reactions, furans, protein modifications, pyrrolysyl tRNA synthetase, X-ray structures

Zitieren

ISO 690

SCHMIDT, Moritz J., Annemarie WEBER, Moritz POTT, Wolfram WELTE, Daniel SUMMERER, 2014. Structural Basis of Furan–Amino Acid Recognition by a Polyspecific Aminoacyl-tRNA-Synthetase and its Genetic Encoding in Human Cells. In: ChemBioChem. 2014, 15(12), pp. 1755-1760. ISSN 1439-4227. eISSN 1439-7633. Available under: doi: 10.1002/cbic.201402006

BibTex

@article{Schmidt2014Struc-29407,
  year={2014},
  doi={10.1002/cbic.201402006},
  title={Structural Basis of Furan–Amino Acid Recognition by a Polyspecific Aminoacyl-tRNA-Synthetase and its Genetic Encoding in Human Cells},
  number={12},
  volume={15},
  issn={1439-4227},
  journal={ChemBioChem},
  pages={1755--1760},
  author={Schmidt, Moritz J. and Weber, Annemarie and Pott, Moritz and Welte, Wolfram and Summerer, Daniel}
}

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