Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin

Mitomycin has a unique chemical structure and contains densely assembled functionalities with extraordinary antitumor activity. The previously proposed mitomycin C biosynthetic pathway has caused great attention to decipher the enzymatic mechanisms for assembling the pharmaceutically unprecedented c...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Dongjin Leng, Yong Sheng, Hengyu Wang, Jianhua Wei, Yixin Ou, Zixin Deng, Linquan Bai, Qianjin Kang
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	mitomycin antitumor acyl carrier protein protein–protein interactions glycosyltransferase reductase Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/f0e03ed512f44f04bf39b96e0ee4fa70
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:f0e03ed512f44f04bf39b96e0ee4fa70
record_format	dspace
spelling	oai:doaj.org-article:f0e03ed512f44f04bf39b96e0ee4fa702021-11-25T18:27:15ZDetermination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin10.3390/molecules262267911420-3049https://doaj.org/article/f0e03ed512f44f04bf39b96e0ee4fa702021-11-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/22/6791https://doaj.org/toc/1420-3049Mitomycin has a unique chemical structure and contains densely assembled functionalities with extraordinary antitumor activity. The previously proposed mitomycin C biosynthetic pathway has caused great attention to decipher the enzymatic mechanisms for assembling the pharmaceutically unprecedented chemical scaffold. Herein, we focused on the determination of acyl carrier protein (ACP)-dependent modification steps and identification of the protein–protein interactions between MmcB (ACP) with the partners in the early-stage biosynthesis of mitomycin C. Based on the initial genetic manipulation consisting of gene disruption and complementation experiments, genes <i>mitE</i>, <i>mmcB</i>, <i>mitB,</i> and <i>mitF</i> were identified as the essential functional genes in the mitomycin C biosynthesis, respectively. Further integration of biochemical analysis elucidated that MitE catalyzed CoA ligation of 3-amino-5-hydroxy-bezonic acid (AHBA), MmcB-tethered AHBA triggered the biosynthesis of mitomycin C, and both MitB and MitF were MmcB-dependent tailoring enzymes involved in the assembly of mitosane. Aiming at understanding the poorly characterized protein–protein interactions, the <i>in vitro</i> pull-down assay was carried out by monitoring MmcB individually with MitB and MitF. The observed results displayed the clear interactions between MmcB and MitB and MitF. The surface plasmon resonance (SPR) biosensor analysis further confirmed the protein–protein interactions of MmcB with MitB and MitF, respectively. Taken together, the current genetic and biochemical analysis will facilitate the investigations of the unusual enzymatic mechanisms for the structurally unique compound assembly and inspire attempts to modify the chemical scaffold of mitomycin family antibiotics.Dongjin LengYong ShengHengyu WangJianhua WeiYixin OuZixin DengLinquan BaiQianjin KangMDPI AGarticlemitomycinantitumoracyl carrier proteinprotein–protein interactionsglycosyltransferasereductaseOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6791, p 6791 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	mitomycin antitumor acyl carrier protein protein–protein interactions glycosyltransferase reductase Organic chemistry QD241-441
spellingShingle	mitomycin antitumor acyl carrier protein protein–protein interactions glycosyltransferase reductase Organic chemistry QD241-441 Dongjin Leng Yong Sheng Hengyu Wang Jianhua Wei Yixin Ou Zixin Deng Linquan Bai Qianjin Kang Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin
description	Mitomycin has a unique chemical structure and contains densely assembled functionalities with extraordinary antitumor activity. The previously proposed mitomycin C biosynthetic pathway has caused great attention to decipher the enzymatic mechanisms for assembling the pharmaceutically unprecedented chemical scaffold. Herein, we focused on the determination of acyl carrier protein (ACP)-dependent modification steps and identification of the protein–protein interactions between MmcB (ACP) with the partners in the early-stage biosynthesis of mitomycin C. Based on the initial genetic manipulation consisting of gene disruption and complementation experiments, genes <i>mitE</i>, <i>mmcB</i>, <i>mitB,</i> and <i>mitF</i> were identified as the essential functional genes in the mitomycin C biosynthesis, respectively. Further integration of biochemical analysis elucidated that MitE catalyzed CoA ligation of 3-amino-5-hydroxy-bezonic acid (AHBA), MmcB-tethered AHBA triggered the biosynthesis of mitomycin C, and both MitB and MitF were MmcB-dependent tailoring enzymes involved in the assembly of mitosane. Aiming at understanding the poorly characterized protein–protein interactions, the <i>in vitro</i> pull-down assay was carried out by monitoring MmcB individually with MitB and MitF. The observed results displayed the clear interactions between MmcB and MitB and MitF. The surface plasmon resonance (SPR) biosensor analysis further confirmed the protein–protein interactions of MmcB with MitB and MitF, respectively. Taken together, the current genetic and biochemical analysis will facilitate the investigations of the unusual enzymatic mechanisms for the structurally unique compound assembly and inspire attempts to modify the chemical scaffold of mitomycin family antibiotics.
format	article
author	Dongjin Leng Yong Sheng Hengyu Wang Jianhua Wei Yixin Ou Zixin Deng Linquan Bai Qianjin Kang
author_facet	Dongjin Leng Yong Sheng Hengyu Wang Jianhua Wei Yixin Ou Zixin Deng Linquan Bai Qianjin Kang
author_sort	Dongjin Leng
title	Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin
title_short	Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin
title_full	Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin
title_fullStr	Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin
title_full_unstemmed	Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin
title_sort	determination of the protein-protein interactions within acyl carrier protein (mmcb)-dependent modifications in the biosynthesis of mitomycin
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f0e03ed512f44f04bf39b96e0ee4fa70
work_keys_str_mv	AT dongjinleng determinationoftheproteinproteininteractionswithinacylcarrierproteinmmcbdependentmodificationsinthebiosynthesisofmitomycin AT yongsheng determinationoftheproteinproteininteractionswithinacylcarrierproteinmmcbdependentmodificationsinthebiosynthesisofmitomycin AT hengyuwang determinationoftheproteinproteininteractionswithinacylcarrierproteinmmcbdependentmodificationsinthebiosynthesisofmitomycin AT jianhuawei determinationoftheproteinproteininteractionswithinacylcarrierproteinmmcbdependentmodificationsinthebiosynthesisofmitomycin AT yixinou determinationoftheproteinproteininteractionswithinacylcarrierproteinmmcbdependentmodificationsinthebiosynthesisofmitomycin AT zixindeng determinationoftheproteinproteininteractionswithinacylcarrierproteinmmcbdependentmodificationsinthebiosynthesisofmitomycin AT linquanbai determinationoftheproteinproteininteractionswithinacylcarrierproteinmmcbdependentmodificationsinthebiosynthesisofmitomycin AT qianjinkang determinationoftheproteinproteininteractionswithinacylcarrierproteinmmcbdependentmodificationsinthebiosynthesisofmitomycin
_version_	1718411170392571904

Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin

Ejemplares similares