Naslov
Discovery and Investigation of Natural Editing Function against Artificial Amino Acids in Protein Translation

Autori
Voller, Jan-Stefan ; Dulić, Morana ; Gerling- Driessen, Ulla I. M. ; Biava, Hernan ; Baumann, Tobias, Budisa, Nediljko ; Gruić-Sovulj, Ita ; Koksch, Beate

Izvornik
ACS central science (2374-7943) 3 (2017); 73-80

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
fluorine, incorporation in proteins, editing, proofreading, aminoacyl-tRNA synthetase, kinetic partitioning

Sažetak
Fluorine being not substantially present in the chemistry of living beings is an attractive element in tailoring novel chemical, biophysical, and pharmacokinetic properties of peptides and proteins. The hallmark of ribosome-mediated artificial amino acid incorporation into peptides and proteins is a broad substrate tolerance, which is assumed to rely on the absence of evolutionary pressure for efficient editing of artificial amino acids. We used the well-characterized editing proficient isoleucyl-tRNA synthetase (IleRS) from Escherichia coli to investigate the crosstalk of aminoacylation and editing activities against fluorinated amino acids. We show that translation of trifluoroethylglycine (TfeGly) into proteins is prevented by hydrolysis of TfeGly-tRNAIle in the IleRS post- transfer editing domain. The remarkable observation is that dissociation of TfeGly- tRNAIle from IleRS is significantly slowed down. This finding is in sharp contrast to natural editing reactions by tRNA synthetases wherein fast editing rates for the noncognate substrates are essential to outcompete fast aa- tRNA dissociation rates. Using a post-transfer editing deficient mutant of IleRS (IleRSAla10), we were able to achieve ribosomal incorporation of TfeGly in vivo. Our work expands the knowledge of ribosome-mediated artificial amino acid translation with detailed analysis of natural editing function against an artificial amino acid providing an impulse for further systematic investigations and engineering of the translation and editing of unusual amino acids.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija

POVEZANOST RADA