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Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site.

B Zhao, L Lei, DG Vassylyev, X Lin, DE Cane, Steven Kelly Orcid Logo, H Yuan, DC Lamb, MR Waterman

Journal of Biological Chemistry, Volume: 284, Issue: 52, Pages: 36711 - 9

Swansea University Author: Steven Kelly Orcid Logo

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DOI (Published version): 10.1074/jbc.M109.064683

Abstract

Albaflavenone synthase (CYP170A1) is a monooxygenase catalyzing the final two steps in the biosynthesis of this antibiotic in the soil bacterium, Streptomyces coelicolor A3(2). Interestingly, CYP170A1 shows no stereo selection forming equal amounts of two albaflavenol epimers, each of which is oxidi...

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Published in: Journal of Biological Chemistry
Published: 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC 2009
URI: https://cronfa.swan.ac.uk/Record/cronfa10330
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spelling 2021-10-29T09:35:46.4384137 v2 10330 2012-03-21 Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site. b17cebaf09b4d737b9378a3581e3de93 0000-0001-7991-5040 Steven Kelly Steven Kelly true false 2012-03-21 BMS Albaflavenone synthase (CYP170A1) is a monooxygenase catalyzing the final two steps in the biosynthesis of this antibiotic in the soil bacterium, Streptomyces coelicolor A3(2). Interestingly, CYP170A1 shows no stereo selection forming equal amounts of two albaflavenol epimers, each of which is oxidized in turn to albaflavenone. To explore the structural basis of the reaction mechanism, we have studied the crystal structures of both ligand-free CYP170A1 (2.6 angstrom) and complex of endogenous substrate (epi-isozizaene) with CYP170A1 (3.3 angstrom). The structure of the complex suggests that the proximal epi-isozizaene molecules may bind to the heme iron in two orientations. In addition, much to our surprise, we have found that albaflavenone synthase also has a second, completely distinct catalytic activity corresponding to the synthesis of farnesene isomers from farnesyl diphosphate. Within the cytochrome P450 alpha-helical domain both the primary sequence and x-ray structure indicate the presence of a novel terpene synthase active site that is moonlighting on the P450 structure. This includes signature sequences for divalent cation binding and an alpha-helical barrel. This barrel is unusual because it consists of only four helices rather than six found in all other terpene synthases. Mutagenesis establishes that this barrel is essential for the terpene synthase activity of CYP170A1 but not for the monooxygenase activity. This is the first bifunctional P450 discovered to have another active site moonlighting on it and the first time a terpene synthase active site is found moonlighting on another protein. Journal Article Journal of Biological Chemistry 284 52 36711 9 AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA STREPTOMYCES-COELICOLOR A3(2); TRANS-BETA-FARNESENE; SESQUITERPENE SYNTHASES; ARISTOLOCHENE SYNTHASE; BIFUNCTIONAL ENZYMES; ALARM PHEROMONE; IDENTIFICATION; BIOSYNTHESIS; CYCLIZATION; VOLATILES 31 12 2009 2009-12-31 10.1074/jbc.M109.064683 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University 2021-10-29T09:35:46.4384137 2012-03-21T16:17:29.0000000 Swansea University Medical School Medicine B Zhao 1 L Lei 2 DG Vassylyev 3 X Lin 4 DE Cane 5 Steven Kelly 0000-0001-7991-5040 6 H Yuan 7 DC Lamb 8 MR Waterman 9
title Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site.
spellingShingle Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site.
Steven Kelly
title_short Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site.
title_full Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site.
title_fullStr Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site.
title_full_unstemmed Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site.
title_sort Crystal structure of albaflavenone monoxygenase containing a moonlighting terpene synthase active site.
author_id_str_mv b17cebaf09b4d737b9378a3581e3de93
author_id_fullname_str_mv b17cebaf09b4d737b9378a3581e3de93_***_Steven Kelly
author Steven Kelly
author2 B Zhao
L Lei
DG Vassylyev
X Lin
DE Cane
Steven Kelly
H Yuan
DC Lamb
MR Waterman
format Journal article
container_title Journal of Biological Chemistry
container_volume 284
container_issue 52
container_start_page 36711
publishDate 2009
institution Swansea University
doi_str_mv 10.1074/jbc.M109.064683
publisher AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
college_str Swansea University Medical School
hierarchytype
hierarchy_top_id swanseauniversitymedicalschool
hierarchy_top_title Swansea University Medical School
hierarchy_parent_id swanseauniversitymedicalschool
hierarchy_parent_title Swansea University Medical School
department_str Medicine{{{_:::_}}}Swansea University Medical School{{{_:::_}}}Medicine
document_store_str 0
active_str 0
description Albaflavenone synthase (CYP170A1) is a monooxygenase catalyzing the final two steps in the biosynthesis of this antibiotic in the soil bacterium, Streptomyces coelicolor A3(2). Interestingly, CYP170A1 shows no stereo selection forming equal amounts of two albaflavenol epimers, each of which is oxidized in turn to albaflavenone. To explore the structural basis of the reaction mechanism, we have studied the crystal structures of both ligand-free CYP170A1 (2.6 angstrom) and complex of endogenous substrate (epi-isozizaene) with CYP170A1 (3.3 angstrom). The structure of the complex suggests that the proximal epi-isozizaene molecules may bind to the heme iron in two orientations. In addition, much to our surprise, we have found that albaflavenone synthase also has a second, completely distinct catalytic activity corresponding to the synthesis of farnesene isomers from farnesyl diphosphate. Within the cytochrome P450 alpha-helical domain both the primary sequence and x-ray structure indicate the presence of a novel terpene synthase active site that is moonlighting on the P450 structure. This includes signature sequences for divalent cation binding and an alpha-helical barrel. This barrel is unusual because it consists of only four helices rather than six found in all other terpene synthases. Mutagenesis establishes that this barrel is essential for the terpene synthase activity of CYP170A1 but not for the monooxygenase activity. This is the first bifunctional P450 discovered to have another active site moonlighting on it and the first time a terpene synthase active site is found moonlighting on another protein.
published_date 2009-12-31T03:19:17Z
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score 10.898474