The Importance of Charge in Perturbing the Aromatic Glue Stabilizing the Protein–Protein Interface of Homodimeric tRNA-Guanine Transglycosylase - DSA-IPHC Accéder directement au contenu
Article Dans Une Revue ACS Chemical Biology Année : 2020

The Importance of Charge in Perturbing the Aromatic Glue Stabilizing the Protein–Protein Interface of Homodimeric tRNA-Guanine Transglycosylase

Résumé

Bacterial tRNA-guanine transglycosylase (Tgt) is involved in the biosynthesis of the modified tRNA nucleoside queuosine present in the anticodon wobble position of tRNAs specific for aspartate, asparagine, histidine and tyrosine. Inactivation of the tgt gene leads to decreased pathogenicity of Shigella bacteria. Therefore, Tgt constitutes a putative target for Shigellosis drug therapy. Since only active as homodimer, interference with dimer-interface formation may, in addition to active-site inhibition, provide further means to disable this protein. A cluster of four aromatic residues seems important to stabilize the homodimer. We mutated residues of this aromatic cluster and analyzed each exchange with respect to dimer and thermal stability or enzyme activity applying native mass spectrometry, thermal shift assay, enzyme kinetics, and X-ray crystallography. Our structural studies indicate strong influence of pH on homodimer stability. Obviously, protonation of a histidine within the aromatic cluster promotes the collapse of an essential structural motif within the dimer interface at slightly acidic pH.
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Dates et versions

hal-03073530 , version 1 (16-12-2020)

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Andreas Nguyen, Dzung Nguyen, Phong Nguyen Tran Xuan, Maurice Sebastiani, Stefanie Dörr, et al.. The Importance of Charge in Perturbing the Aromatic Glue Stabilizing the Protein–Protein Interface of Homodimeric tRNA-Guanine Transglycosylase. ACS Chemical Biology, 2020, 15 (11), pp.3021-3029. ⟨10.1021/acschembio.0c00700⟩. ⟨hal-03073530⟩
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