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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Autores principales:	Chang-Sook Jeong, Jisub Hwang, Hackwon Do, Sun-Shin Cha, Tae-Jin Oh, Hak Jun Kim, Hyun Ho Park, Jun Hyuck Lee
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/263a99af7e764dfaa8e2975b4b8abcdc
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spelling	oai:doaj.org-article:263a99af7e764dfaa8e2975b4b8abcdc2021-12-02T15:11:52ZStructural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis10.1038/s41598-020-78699-z2045-2322https://doaj.org/article/263a99af7e764dfaa8e2975b4b8abcdc2020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78699-zhttps://doaj.org/toc/2045-2322Abstract 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.Chang-Sook JeongJisub HwangHackwon DoSun-Shin ChaTae-Jin OhHak Jun KimHyun Ho ParkJun Hyuck LeeNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-14 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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
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.
format	article
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
title	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_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_sort	structural and biochemical analyses of an aminoglycoside 2′-n-acetyltransferase from mycolicibacterium smegmatis
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/263a99af7e764dfaa8e2975b4b8abcdc
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Structural and biochemical analyses of an aminoglycoside 2′-N-acetyltransferase from Mycolicibacterium smegmatis

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