4CS2

Catalytic domain of Pyrrolysyl-tRNA synthetase mutant Y306A, Y384F in its apo form

Summary for 4CS2

• Entry DOI: 10.2210/pdb4cs2/pdb
• Descriptor: PYRROLYSINE--TRNA LIGASE, DI(HYDROXYETHYL)ETHER, 1,2-ETHANEDIOL, ... (4 entities in total)
• Functional Keywords: ligase
• Biological source: METHANOSARCINA MAZEI
• Cellular location: Cytoplasm (By similarity): Q8PWY1
• Total number of polymer chains: 1
• Total formula weight: 32090.80

Authors

Schmidt, M.J.,Weber, A.,Pott, M.,Welte, W.,Summerer, D. (deposition date: 2014-03-04, release date: 2014-04-30, Last modification date: 2023-12-20)

Primary citation

Schmidt, M.J.,Weber, A.,Pott, M.,Welte, W.,Summerer, D.
Structural Basis of Furan-Amino Acid Recognition by a Polyspecific Aminoacyl-tRNA-Synthetase and its Genetic Encoding in Human Cells.
Chembiochem, 15:1755-, 2014

PubMed Abstract: 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.

PubMed: 24737732
DOI: 10.1002/CBIC.201402006

Experimental method

X-RAY DIFFRACTION (1.9 Å)