Isolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer

ABSTRACT The integral membrane protein heme A synthase (HAS) catalyzes the biosynthesis of heme A, which is a prerequisite for cellular respiration in a wide range of aerobic organisms. Previous studies have revealed that HAS can form homo-oligomeric complexes, and this oligomerization appears to be...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Hui Zeng, Guoliang Zhu, Shuangbo Zhang, Xinmei Li, Janosch Martin, Nina Morgner, Fei Sun, Guohong Peng, Hao Xie, Hartmut Michel
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Aquifex aeolicus cofactor biosynthesis heme A synthase hyperthermophilic bacterium metalloproteins protein oligomerization Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/373e80c3052d423d82f3ea674c8bff61
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:373e80c3052d423d82f3ea674c8bff61
record_format	dspace
spelling	oai:doaj.org-article:373e80c3052d423d82f3ea674c8bff612021-11-15T15:56:47ZIsolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer10.1128/mBio.02615-192150-7511https://doaj.org/article/373e80c3052d423d82f3ea674c8bff612020-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02615-19https://doaj.org/toc/2150-7511ABSTRACT The integral membrane protein heme A synthase (HAS) catalyzes the biosynthesis of heme A, which is a prerequisite for cellular respiration in a wide range of aerobic organisms. Previous studies have revealed that HAS can form homo-oligomeric complexes, and this oligomerization appears to be evolutionarily conserved among prokaryotes and eukaryotes and is shown to be essential for the biological function of eukaryotic HAS. Despite its importance, little is known about the detailed structural properties of HAS oligomers. Here, we aimed to address this critical issue by analyzing the oligomeric state of HAS from Aquifex aeolicus (AaHAS) using a combination of techniques, including size exclusion chromatography coupled with multiangle light scattering (SEC-MALS), cross-linking, laser-induced liquid bead ion desorption mass spectrometry (LILBID-MS), and single-particle electron cryomicroscopy (cryo-EM). Our results show that HAS forms a thermostable trimeric complex. A cryo-EM density map provides information on the oligomerization interface of the AaHAS trimer. These results provide structural insights into HAS multimerization and expand our knowledge of this important enzyme. IMPORTANCE Heme A is a vital redox cofactor unique for the terminal cytochrome c oxidase in mitochondria and many microorganisms. It plays a key role in oxygen reduction by serving as an electron carrier and as the oxygen-binding site. Heme A is synthesized from heme O by an integral membrane protein, heme A synthase (HAS). Defects in HAS impair cellular respiration and have been linked to various human diseases, e.g., fatal infantile hypertrophic cardiomyopathy and Leigh syndrome. HAS exists as a stable oligomeric complex, and studies have shown that oligomerization of eukaryotic HAS is necessary for its proper function. However, the molecular architecture of the HAS oligomeric complex has remained uncharacterized. The present study shows that HAS forms trimers and reveals how the oligomeric arrangement contributes to the complex stability and flexibility, enabling HAS to perform its catalytic function effectively. This work provides the basic understanding for future studies on heme A biosynthesis.Hui ZengGuoliang ZhuShuangbo ZhangXinmei LiJanosch MartinNina MorgnerFei SunGuohong PengHao XieHartmut MichelAmerican Society for MicrobiologyarticleAquifex aeolicuscofactor biosynthesisheme A synthasehyperthermophilic bacteriummetalloproteinsprotein oligomerizationMicrobiologyQR1-502ENmBio, Vol 11, Iss 3 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Aquifex aeolicus cofactor biosynthesis heme A synthase hyperthermophilic bacterium metalloproteins protein oligomerization Microbiology QR1-502
spellingShingle	Aquifex aeolicus cofactor biosynthesis heme A synthase hyperthermophilic bacterium metalloproteins protein oligomerization Microbiology QR1-502 Hui Zeng Guoliang Zhu Shuangbo Zhang Xinmei Li Janosch Martin Nina Morgner Fei Sun Guohong Peng Hao Xie Hartmut Michel Isolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer
description	ABSTRACT The integral membrane protein heme A synthase (HAS) catalyzes the biosynthesis of heme A, which is a prerequisite for cellular respiration in a wide range of aerobic organisms. Previous studies have revealed that HAS can form homo-oligomeric complexes, and this oligomerization appears to be evolutionarily conserved among prokaryotes and eukaryotes and is shown to be essential for the biological function of eukaryotic HAS. Despite its importance, little is known about the detailed structural properties of HAS oligomers. Here, we aimed to address this critical issue by analyzing the oligomeric state of HAS from Aquifex aeolicus (AaHAS) using a combination of techniques, including size exclusion chromatography coupled with multiangle light scattering (SEC-MALS), cross-linking, laser-induced liquid bead ion desorption mass spectrometry (LILBID-MS), and single-particle electron cryomicroscopy (cryo-EM). Our results show that HAS forms a thermostable trimeric complex. A cryo-EM density map provides information on the oligomerization interface of the AaHAS trimer. These results provide structural insights into HAS multimerization and expand our knowledge of this important enzyme. IMPORTANCE Heme A is a vital redox cofactor unique for the terminal cytochrome c oxidase in mitochondria and many microorganisms. It plays a key role in oxygen reduction by serving as an electron carrier and as the oxygen-binding site. Heme A is synthesized from heme O by an integral membrane protein, heme A synthase (HAS). Defects in HAS impair cellular respiration and have been linked to various human diseases, e.g., fatal infantile hypertrophic cardiomyopathy and Leigh syndrome. HAS exists as a stable oligomeric complex, and studies have shown that oligomerization of eukaryotic HAS is necessary for its proper function. However, the molecular architecture of the HAS oligomeric complex has remained uncharacterized. The present study shows that HAS forms trimers and reveals how the oligomeric arrangement contributes to the complex stability and flexibility, enabling HAS to perform its catalytic function effectively. This work provides the basic understanding for future studies on heme A biosynthesis.
format	article
author	Hui Zeng Guoliang Zhu Shuangbo Zhang Xinmei Li Janosch Martin Nina Morgner Fei Sun Guohong Peng Hao Xie Hartmut Michel
author_facet	Hui Zeng Guoliang Zhu Shuangbo Zhang Xinmei Li Janosch Martin Nina Morgner Fei Sun Guohong Peng Hao Xie Hartmut Michel
author_sort	Hui Zeng
title	Isolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer
title_short	Isolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer
title_full	Isolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer
title_fullStr	Isolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer
title_full_unstemmed	Isolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer
title_sort	isolated heme a synthase from <named-content content-type="genus-species">aquifex aeolicus</named-content> is a trimer
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/373e80c3052d423d82f3ea674c8bff61
work_keys_str_mv	AT huizeng isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT guoliangzhu isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT shuangbozhang isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT xinmeili isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT janoschmartin isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT ninamorgner isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT feisun isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT guohongpeng isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT haoxie isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer AT hartmutmichel isolatedhemeasynthasefromnamedcontentcontenttypegenusspeciesaquifexaeolicusnamedcontentisatrimer
_version_	1718427079577436160

Isolated Heme A Synthase from <named-content content-type="genus-species">Aquifex aeolicus</named-content> Is a Trimer

Ejemplares similares