Molecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)

ABSTRACT The underlying molecular mechanisms of flavin-dependent amine oxidases remain relatively poorly understood, even though many of these enzymes have been reported. The nicotine oxidoreductase NicA2 is a crucial enzyme for the first step of nicotine degradation in Pseudomonas putida S16 (DSM 2...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Hongzhi Tang, Kunzhi Zhang, Haiyang Hu, Geng Wu, Weiwei Wang, Xiongyu Zhu, Gongquan Liu, Ping Xu
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	nicotine oxidoreductase NicA2 pseudooxynicotine crystal structure molecular mechanism Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/648baaf785bc4c5fb0089120acde63fb
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:648baaf785bc4c5fb0089120acde63fb
record_format	dspace
spelling	oai:doaj.org-article:648baaf785bc4c5fb0089120acde63fb2021-11-15T16:19:09ZMolecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)10.1128/mBio.02012-202150-7511https://doaj.org/article/648baaf785bc4c5fb0089120acde63fb2020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02012-20https://doaj.org/toc/2150-7511ABSTRACT The underlying molecular mechanisms of flavin-dependent amine oxidases remain relatively poorly understood, even though many of these enzymes have been reported. The nicotine oxidoreductase NicA2 is a crucial enzyme for the first step of nicotine degradation in Pseudomonas putida S16 (DSM 28022). Here, we present the crystal structure of a ternary complex comprising NicA2 residues 21 to 482, flavin adenine dinucleotide (FAD), and nicotine at 2.25 Å resolution. Unlike other, related structures, NicA2 does not have an associated diacyl glycerophospholipid, wraps its substrate more tightly, and has an intriguing exit passage in which nine bulky amino acid residues occlude the release of its toxic product, pseudooxynicotine (PN). The replacement of these bulky residues by amino acids with small side chains effectively increases the catalytic turnover rate of NicA2. Our results indicate that the passage in wild-type NicA2 effectively controls the rate of PN release and thus prevents its rapid intracellular accumulation. It gives ample time for PN to be converted to less-harmful substances by downstream enzymes such as pseudooxynicotine amine oxidase (Pnao) before its accumulation causes cell damage or even death. The temporal metabolic regulation mode revealed in this study may shed light on the production of cytotoxic compounds. IMPORTANCE Flavin-dependent amine oxidases have received extensive attention because of their importance in drug metabolism, Parkinson’s disease, and neurotransmitter catabolism. However, the underlying molecular mechanisms remain relatively poorly understood. Here, combining the crystal structure of NicA2 (an enzyme in the first step of the bacterial nicotine degradation pathway in Pseudomonas putida S16 (DSM 28022)), biochemical analysis, and mutant construction, we found an intriguing exit passage in which bulky amino acid residues occlude the release of the toxic product of NicA2, in contrast to other, related structures. The selective product exportation register for NicA2 has proven to be beneficial to cell growth. Those seeking to produce cytotoxic compounds could greatly benefit from the use of such an export register mechanism.Hongzhi TangKunzhi ZhangHaiyang HuGeng WuWeiwei WangXiongyu ZhuGongquan LiuPing XuAmerican Society for Microbiologyarticlenicotine oxidoreductaseNicA2pseudooxynicotinecrystal structuremolecular mechanismMicrobiologyQR1-502ENmBio, Vol 11, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	nicotine oxidoreductase NicA2 pseudooxynicotine crystal structure molecular mechanism Microbiology QR1-502
spellingShingle	nicotine oxidoreductase NicA2 pseudooxynicotine crystal structure molecular mechanism Microbiology QR1-502 Hongzhi Tang Kunzhi Zhang Haiyang Hu Geng Wu Weiwei Wang Xiongyu Zhu Gongquan Liu Ping Xu Molecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)
description	ABSTRACT The underlying molecular mechanisms of flavin-dependent amine oxidases remain relatively poorly understood, even though many of these enzymes have been reported. The nicotine oxidoreductase NicA2 is a crucial enzyme for the first step of nicotine degradation in Pseudomonas putida S16 (DSM 28022). Here, we present the crystal structure of a ternary complex comprising NicA2 residues 21 to 482, flavin adenine dinucleotide (FAD), and nicotine at 2.25 Å resolution. Unlike other, related structures, NicA2 does not have an associated diacyl glycerophospholipid, wraps its substrate more tightly, and has an intriguing exit passage in which nine bulky amino acid residues occlude the release of its toxic product, pseudooxynicotine (PN). The replacement of these bulky residues by amino acids with small side chains effectively increases the catalytic turnover rate of NicA2. Our results indicate that the passage in wild-type NicA2 effectively controls the rate of PN release and thus prevents its rapid intracellular accumulation. It gives ample time for PN to be converted to less-harmful substances by downstream enzymes such as pseudooxynicotine amine oxidase (Pnao) before its accumulation causes cell damage or even death. The temporal metabolic regulation mode revealed in this study may shed light on the production of cytotoxic compounds. IMPORTANCE Flavin-dependent amine oxidases have received extensive attention because of their importance in drug metabolism, Parkinson’s disease, and neurotransmitter catabolism. However, the underlying molecular mechanisms remain relatively poorly understood. Here, combining the crystal structure of NicA2 (an enzyme in the first step of the bacterial nicotine degradation pathway in Pseudomonas putida S16 (DSM 28022)), biochemical analysis, and mutant construction, we found an intriguing exit passage in which bulky amino acid residues occlude the release of the toxic product of NicA2, in contrast to other, related structures. The selective product exportation register for NicA2 has proven to be beneficial to cell growth. Those seeking to produce cytotoxic compounds could greatly benefit from the use of such an export register mechanism.
format	article
author	Hongzhi Tang Kunzhi Zhang Haiyang Hu Geng Wu Weiwei Wang Xiongyu Zhu Gongquan Liu Ping Xu
author_facet	Hongzhi Tang Kunzhi Zhang Haiyang Hu Geng Wu Weiwei Wang Xiongyu Zhu Gongquan Liu Ping Xu
author_sort	Hongzhi Tang
title	Molecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)
title_short	Molecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)
title_full	Molecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)
title_fullStr	Molecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)
title_full_unstemmed	Molecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)
title_sort	molecular deceleration regulates toxicant release to prevent cell damage in <named-content content-type="genus-species">pseudomonas putida</named-content> s16 (dsm 28022)
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/648baaf785bc4c5fb0089120acde63fb
work_keys_str_mv	AT hongzhitang moleculardecelerationregulatestoxicantreleasetopreventcelldamageinnamedcontentcontenttypegenusspeciespseudomonasputidanamedcontents16dsm28022 AT kunzhizhang moleculardecelerationregulatestoxicantreleasetopreventcelldamageinnamedcontentcontenttypegenusspeciespseudomonasputidanamedcontents16dsm28022 AT haiyanghu moleculardecelerationregulatestoxicantreleasetopreventcelldamageinnamedcontentcontenttypegenusspeciespseudomonasputidanamedcontents16dsm28022 AT gengwu moleculardecelerationregulatestoxicantreleasetopreventcelldamageinnamedcontentcontenttypegenusspeciespseudomonasputidanamedcontents16dsm28022 AT weiweiwang moleculardecelerationregulatestoxicantreleasetopreventcelldamageinnamedcontentcontenttypegenusspeciespseudomonasputidanamedcontents16dsm28022 AT xiongyuzhu moleculardecelerationregulatestoxicantreleasetopreventcelldamageinnamedcontentcontenttypegenusspeciespseudomonasputidanamedcontents16dsm28022 AT gongquanliu moleculardecelerationregulatestoxicantreleasetopreventcelldamageinnamedcontentcontenttypegenusspeciespseudomonasputidanamedcontents16dsm28022 AT pingxu moleculardecelerationregulatestoxicantreleasetopreventcelldamageinnamedcontentcontenttypegenusspeciespseudomonasputidanamedcontents16dsm28022
_version_	1718426904501944320

Molecular Deceleration Regulates Toxicant Release to Prevent Cell Damage in <named-content content-type="genus-species">Pseudomonas putida</named-content> S16 (DSM 28022)

Ejemplares similares