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Structural and biochemical analyses of an aminoglycoside 2?-N-acetyltransferase from Mycolicibacterium smegmatis
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View all Nature Research journals Search My Account Loginnature scientific reports articles article Structural and biochemical analyses of an aminoglycoside 2?-N-acetyltransferase from Mycolicibacterium smegmatis Download PDF
Subjects AbstractThe expression of aminoglycoside-modifying enzymes represents a survival strategy of antibiotic-resistant bacteria. Aminoglycoside 2?-N-acetyltransferase [AAC(2?)] neutralizes aminoglycoside drugs by acetylation of their 2? amino groups in an acetyl coenzyme A (CoA)-dependent manner. To understand the structural features and molecular mechanism underlying AAC(2?) activity, we overexpressed, purified, and crystallized AAC(2?) from Mycolicibacterium smegmatis [AAC(2?)-Id] and determined the crystal structures of its apo-form and ternary complexes with CoA and four different aminoglycosides (gentamicin, sisomicin, neomycin, and paromomycin). These AAC(2?)-Id structures unraveled the binding modes of different aminoglycosides, explaining the broad substrate specificity of the enzyme. Comparative structural analysis showed that the ?4-helix and ?8–?9 loop region undergo major conformational changes upon CoA and substrate binding. Additionally, structural comparison between the present paromomycin-bound AAC(2?)-Id structure and the previously reported paromomycin-bound AAC(6?)-Ib and 30S ribosome structures revealed the structural features of paromomycin that are responsible for its antibiotic activity and AAC binding. Taken together, these results provide useful information for designing AAC(2?) inhibitors and for the chemical modification of aminoglycosides.
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- Article
- Open Access
- Published: 09 December 2020
- Chang-Sook Jeong1,2,
- Jisub Hwang1,2,
- Hackwon Do1,
- Sun-Shin Cha3,
- Tae-Jin Oh4,5,6,
- Hak Jun Kim7,
- Hyun Ho Park8 &
- Jun Hyuck Lee1,2
Subjects AbstractThe expression of aminoglycoside-modifying enzymes represents a survival strategy of antibiotic-resistant bacteria. Aminoglycoside 2?-N-acetyltransferase [AAC(2?)] neutralizes aminoglycoside drugs by acetylation of their 2? amino groups in an acetyl coenzyme A (CoA)-dependent manner. To understand the structural features and molecular mechanism underlying AAC(2?) activity, we overexpressed, purified, and crystallized AAC(2?) from Mycolicibacterium smegmatis [AAC(2?)-Id] and determined the crystal structures of its apo-form and ternary complexes with CoA and four different aminoglycosides (gentamicin, sisomicin, neomycin, and paromomycin). These AAC(2?)-Id structures unraveled the binding modes of different aminoglycosides, explaining the broad substrate specificity of the enzyme. Comparative structural analysis showed that the ?4-helix and ?8–?9 loop region undergo major conformational changes upon CoA and substrate binding. Additionally, structural comparison between the present paromomycin-bound AAC(2?)-Id structure and the previously reported paromomycin-bound AAC(6?)-Ib and 30S ribosome structures revealed the structural features of paromomycin that are responsible for its antibiotic activity and AAC binding. Taken together, these results provide useful information for designing AAC(2?) inhibitors and for the chemical modification of aminoglycosides.
Publication date: 09/12/2020
Author: Chang-Sook Jeong
Reference: doi:10.1038/s41598-020-78699-z
This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 870292.
