PXD048763 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Bacterial stigmasterol degradation involving radical flavin delta-24 desaturase and molybdenum-dependent C26 hydroxylase |
Description | Sterols are ubiquitous membrane constituents that persist to a large extent in the environment due to their water insolubility and chemical inertness. Recently, an oxygenase-independent sterol degradation pathway was discovered in a cholesterol-grown denitrifying bacterium Sterolibacterium (S.) denitrificans. It achieves hydroxylation of the unactivated primary C26 of the isoprenoid side chain to an allylic alcohol via a phosphorylated intermediate in a four-step ATP-dependent enzyme cascade. However, this pathway is incompatible with the degradation of widely distributed steroids containing a double bond at C-22 in the isoprenoid side chain such as the plant sterol stigmasterol. Here, we have enriched a prototypical delta-24 desaturase from S. denitrificans, which catalyses the electron acceptor-dependent oxidation of the intermediate stigmast-1,4-diene-3-one (SDO) to a conjugated 22,(E)24-diene. We suggest an 44 architecture of the 440 kDa enzyme, with each subunit covalently binding an FMN cofactor to a histidyl residue. As isolated, both flavins are present as red semiquinone radicals, which can be reduced by SDO but cannot be oxidised even with strong oxidising agents. We propose a mechanism involving an allylic radical intermediate in which two flavin semiquinones each abstract one hydrogen atom from the substrate. The conjugated delta-22,24 moiety formed allows for the subsequent hydroxylation of the terminal C26 with water by a heterologously produced molybdenum-dependent steroid C26 dehydrogenase. In conclusion, the pathway elucidated for delta-22 steroids achieves oxygen-independent hydroxylation of the isoprenoid side chain by bypassing the ATP-dependent formation of a phosphorylated intermediate. |
HostingRepository | PRIDE |
AnnounceDate | 2024-10-22 |
AnnouncementXML | Submission_2024-10-22_06:39:50.041.xml |
DigitalObjectIdentifier | https://dx.doi.org/10.6019/PXD048763 |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Supported dataset by repository |
PrimarySubmitter | Friedel Drepper |
SpeciesList | scientific name: Sterolibacterium denitrificans; NCBI TaxID: 157592; |
ModificationList | 1'-(8alpha-FMN)-L-histidine |
Instrument | Q Exactive Plus |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2024-01-22 06:17:04 | ID requested | |
1 | 2024-05-21 06:53:24 | announced | |
⏵ 2 | 2024-10-22 06:39:54 | announced | 2024-10-22: Updated project metadata. |
Publication List
10.1016/j.jbc.2024.107243; |
10.6019/PXD048763; |
Zhan T, Jacoby C, Jede M, Knapp B, Ferlaino S, G, ü, nter A, Drepper F, M, ü, ller M, Weber S, Boll M, Bacterial stigmasterol degradation involving radical flavin delta-24 desaturase and molybdenum-dependent C26 hydroxylase. J Biol Chem, 300(5):107243(2024) [pubmed] |
Keyword List
submitter keyword: anaerobic metabolism, molybdenum enzyme,steroid degradation, flavin radical enzyme, stigmasterol, desaturasem |
Contact List
Friedel Drepper |
contact affiliation | University of Freiburg, Institute of Biology II, Schänzlestr. 1, 79104 Freiburg, Germany |
contact email | f.drepper@biologie.uni-freiburg.de |
lab head | |
Friedel Drepper |
contact affiliation | University of Freiburg |
contact email | f.drepper@biologie.uni-freiburg.de |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD048763
- Label: PRIDE project
- Name: Bacterial stigmasterol degradation involving radical flavin delta-24 desaturase and molybdenum-dependent C26 hydroxylase