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Article Dans Une Revue Journal of Structural Biology Année : 2020

Use of β3-methionine as an amino acid substrate of Escherichia coli methionyl-tRNA synthetase

Résumé

Polypeptides containing β-amino acids are attractive tools for the design of novel proteins having unique properties of medical or industrial interest. Incorporation of β-amino acids in vivo requires the development of efficient aminoacyl-tRNA synthetases specific of these non-canonical amino acids. Here, we have performed a detailed structural and biochemical study of the recognition and use of β3-Met by Escherichia coli methionyl-tRNA synthetase (MetRS). We show that MetRS binds β3-Met with a 24-fold lower affinity but catalyzes the esterification of the non-canonical amino acid onto tRNA with a rate lowered by three orders of magnitude. Accurate measurements of the catalytic parameters required careful consideration of the presence of contaminating α-Met in β3-Met commercial samples. The 1.45 Å crystal structure of the MetRS: β3-Met complex shows that β3-Met binds the enzyme essentially like α-Met, but the carboxylate moiety is mobile and not adequately positioned to react with ATP for aminoacyl adenylate formation. This study provides structural and biochemical bases for engineering MetRS with improved β3-Met aminoacylation capabilities.
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Origine : Publication financée par une institution

Dates et versions

hal-02493753 , version 1 (26-11-2020)

Identifiants

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Giuliano Nigro, Sophie Bourcier, Christine Lazennec-Schurdevin, Emmanuelle Schmitt, Philippe Marlière, et al.. Use of β3-methionine as an amino acid substrate of Escherichia coli methionyl-tRNA synthetase. Journal of Structural Biology, 2020, 209 (2), pp.107435. ⟨10.1016/j.jsb.2019.107435⟩. ⟨hal-02493753⟩
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