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Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase

Aminoglycoside resistance is commonly conferred by enzymatic modification of drugs by aminoglycoside-modifying enzymes such as aminoglycoside nucleo­tidyltransferases (ANTs). Here, the first crystal structure of an ANT(3′′)(9) adenyltransferase, AadA from Salmonella enterica, is presented. AadA cata...

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Autores principales: Chen, Yang, Näsvall, Joakim, Wu, Shiying, Andersson, Dan I., Selmer, Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4631478/
https://www.ncbi.nlm.nih.gov/pubmed/26527143
http://dx.doi.org/10.1107/S1399004715016429
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author Chen, Yang
Näsvall, Joakim
Wu, Shiying
Andersson, Dan I.
Selmer, Maria
author_facet Chen, Yang
Näsvall, Joakim
Wu, Shiying
Andersson, Dan I.
Selmer, Maria
author_sort Chen, Yang
collection PubMed
description Aminoglycoside resistance is commonly conferred by enzymatic modification of drugs by aminoglycoside-modifying enzymes such as aminoglycoside nucleo­tidyltransferases (ANTs). Here, the first crystal structure of an ANT(3′′)(9) adenyltransferase, AadA from Salmonella enterica, is presented. AadA catalyses the magnesium-dependent transfer of adenosine monophosphate from ATP to the two chemically dissimilar drugs streptomycin and spectinomycin. The structure was solved using selenium SAD phasing and refined to 2.5 Å resolution. AadA consists of a nucleotidyltransferase domain and an α-helical bundle domain. AadA crystallizes as a monomer and is a monomer in solution as confirmed by small-angle X-ray scattering, in contrast to structurally similar homodimeric adenylating enzymes such as kanamycin nucleotidyltransferase. Isothermal titration calorimetry experiments show that ATP binding has to occur before binding of the aminoglycoside substrate, and structure analysis suggests that ATP binding repositions the two domains for aminoglycoside binding in the interdomain cleft. Candidate residues for ligand binding and catalysis were subjected to site-directed mutagenesis. In vivo resistance and in vitro binding assays support the role of Glu87 as the catalytic base in adenylation, while Arg192 and Lys205 are shown to be critical for ATP binding.
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spelling pubmed-46314782015-11-20 Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase Chen, Yang Näsvall, Joakim Wu, Shiying Andersson, Dan I. Selmer, Maria Acta Crystallogr D Biol Crystallogr Research Papers Aminoglycoside resistance is commonly conferred by enzymatic modification of drugs by aminoglycoside-modifying enzymes such as aminoglycoside nucleo­tidyltransferases (ANTs). Here, the first crystal structure of an ANT(3′′)(9) adenyltransferase, AadA from Salmonella enterica, is presented. AadA catalyses the magnesium-dependent transfer of adenosine monophosphate from ATP to the two chemically dissimilar drugs streptomycin and spectinomycin. The structure was solved using selenium SAD phasing and refined to 2.5 Å resolution. AadA consists of a nucleotidyltransferase domain and an α-helical bundle domain. AadA crystallizes as a monomer and is a monomer in solution as confirmed by small-angle X-ray scattering, in contrast to structurally similar homodimeric adenylating enzymes such as kanamycin nucleotidyltransferase. Isothermal titration calorimetry experiments show that ATP binding has to occur before binding of the aminoglycoside substrate, and structure analysis suggests that ATP binding repositions the two domains for aminoglycoside binding in the interdomain cleft. Candidate residues for ligand binding and catalysis were subjected to site-directed mutagenesis. In vivo resistance and in vitro binding assays support the role of Glu87 as the catalytic base in adenylation, while Arg192 and Lys205 are shown to be critical for ATP binding. International Union of Crystallography 2015-10-31 /pmc/articles/PMC4631478/ /pubmed/26527143 http://dx.doi.org/10.1107/S1399004715016429 Text en © Chen et al. 2015 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Research Papers
Chen, Yang
Näsvall, Joakim
Wu, Shiying
Andersson, Dan I.
Selmer, Maria
Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase
title Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase
title_full Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase
title_fullStr Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase
title_full_unstemmed Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase
title_short Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase
title_sort structure of aada from salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4631478/
https://www.ncbi.nlm.nih.gov/pubmed/26527143
http://dx.doi.org/10.1107/S1399004715016429
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