Structural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis
Abstract The 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 und...
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Nature Portfolio
2020-12-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-020-78699-z |
| _version_ | 1818346965716434944 |
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| author | Chang-Sook Jeong Jisub Hwang Hackwon Do Sun-Shin Cha Tae-Jin Oh Hak Jun Kim Hyun Ho Park Jun Hyuck Lee |
| author_facet | Chang-Sook Jeong Jisub Hwang Hackwon Do Sun-Shin Cha Tae-Jin Oh Hak Jun Kim Hyun Ho Park Jun Hyuck Lee |
| author_sort | Chang-Sook Jeong |
| collection | DOAJ |
| description | Abstract The 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 α4-helix and β8–β9 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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| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-13T17:26:39Z |
| publishDate | 2020-12-01 |
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| series | Scientific Reports |
| spelling | doaj.art-263a99af7e764dfaa8e2975b4b8abcdc2022-12-21T23:37:10ZengNature PortfolioScientific Reports2045-23222020-12-0110111410.1038/s41598-020-78699-zStructural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatisChang-Sook Jeong0Jisub Hwang1Hackwon Do2Sun-Shin Cha3Tae-Jin Oh4Hak Jun Kim5Hyun Ho Park6Jun Hyuck Lee7Research Unit of Cryogenic Novel Material, Korea Polar Research InstituteResearch Unit of Cryogenic Novel Material, Korea Polar Research InstituteResearch Unit of Cryogenic Novel Material, Korea Polar Research InstituteDepartment of Chemistry and Nanoscience, Ewha Womans UniversityDepartment of Life Science and Biochemical Engineering, Graduate School, SunMoon UniversityDepartment of Chemistry, Pukyong National UniversityCollege of Pharmacy, Chung-Ang UniversityResearch Unit of Cryogenic Novel Material, Korea Polar Research InstituteAbstract The 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 α4-helix and β8–β9 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.https://doi.org/10.1038/s41598-020-78699-z |
| spellingShingle | Chang-Sook Jeong Jisub Hwang Hackwon Do Sun-Shin Cha Tae-Jin Oh Hak Jun Kim Hyun Ho Park Jun Hyuck Lee Structural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis Scientific Reports |
| title | Structural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis |
| title_full | Structural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis |
| title_fullStr | Structural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis |
| title_full_unstemmed | Structural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis |
| title_short | Structural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis |
| title_sort | structural and biochemical analyses of an aminoglycoside 2 n acetyltransferase from mycolicibacterium smegmatis |
| url | https://doi.org/10.1038/s41598-020-78699-z |
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