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Investigating the conformational changes of OEA on HIF-3A-ARNT by HDX-MS

ABSTRACT: We used hydrogen-deuterium exchange mass spectrometry (HDX-MS) to investigate the influence of OEA on HIF-3A-ARNT.HDX-MS data indicated that OEA binding stabilized multiple residues within the Gβ and Fα regions of HIF-3A PAS-B domain.It is possible that the stabilized Gβ and Fα function as an “interlayer” to further contact with the AB-loop of ARNT.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Xiaotong Diao

LAB HEAD: Dalei Wu

PROVIDER: PXD033376 | Pride | 2022-05-20

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	HIF3-OEA.hdx	Other
	HIF3_FullD_fullMS_D2O_0s_0_20200829_201226.raw	Raw
	HIF3_FullD_fullMS_D2O_0s_0_20200901_114356.raw	Raw
	HIF3_OEA_1_fullMS_D2O_0s_0_20200828_193031.raw	Raw
	HIF3_OEA_1_fullMS_D2O_10000s_10000_20200829_011949.raw	Raw

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Publications

Identification of oleoylethanolamide as an endogenous ligand for HIF-3α.

Diao Xiaotong X Ye Fei F Zhang Meina M Ren Xintong X Tian Xiaoxu X Lu Jingping J Sun Xiangnan X Hou Zeng Z Chen Xiaoyu X Li Fengwei F Zhuang Jingjing J Ding Hong H Peng Chao C Rastinejad Fraydoon F Luo Cheng C Wu Dalei D

Nature communications 20220509 1

Hypoxia-inducible factors (HIFs) are α/β heterodimeric transcription factors modulating cellular responses to the low oxygen condition. Among three HIF-α isoforms, HIF-3α is the least studied to date. Here we show that oleoylethanolamide (OEA), a physiological lipid known to regulate food intake and metabolism, binds selectively to HIF-3α. Through crystallographic analysis of HIF-3 α/β heterodimer in both apo and OEA-bound forms, hydrogen-deuterium exchange mass spectrometry (HDX-MS), molecular ...[more]

PMID: 35534502

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2023-06-19 | PXD042971 | Pride

HDX-MS on secondary transporters

Project description:Secondary transporters undergo structural rearrangements to catalyze substrate translocation across the cell membrane – yet how such conformational changes happen within a lipid environment remains poorly understood. Here, we combine hydrogen-deuterium exchange mass spectrometry (HDX-MS) with molecular dynamics (MD) simulations to understand how lipids regulate the conformational dynamics of secondary transporters at the molecular level. Using the homologous transporters XylE, LacY and GlpT from Escherichia coli as model systems, we discover that conserved networks of charged residues act as molecular switches that drive the conformational transition between different states. We reveal that these molecular switches are regulated by interactions with surrounding phospholipids and show that phosphatidylethanolamine interferes with the formation of the conserved networks and favors an inward-facing state. Overall, this work provides insights into the importance of lipids in shaping the conformational landscape of an important class of transporters.

2018-10-16 | PXD011060 | Pride

Structural basis for ALK2/BMPR2 receptor complex signaling through kinase domain oligomerization

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2021-08-16 | PXD022944 | Pride

HDX-MS of CSN-CRL2 complexes - Structural basis of Cullin 2 RING E3 ligase regulation by the COP9 signalosome

Project description:Cullin-Ring E3 Ligases (CRLs) regulate a multitude of cellular pathways through specific substrate receptors. The COP9 signalosome (CSN) deactivates CRLs by removing NEDD8 (N8) from activated Cullins. The structure of stable CSN-CRL can be used to understand this mechanism of regulation. Here we present the first structures of the neddylated and deneddylated CSN-CRL2 complexes by combining single particle cryo-electron microscopy (cryo-EM) with chemical cross-linking mass spectrometry (MS). These structures reveal a conserved mechanism of CSN activation, consisting of conformational clamping of the CRL2 substrate by CSN2/CSN4, release of the catalytic CSN5/CSN6 heterodimer and finally activation of the CSN5 deneddylation machinery. Using hydrogen deuterium exchange-MS we show that CRL2 binding and conformational activation of CSN5/CSN6 occur in a neddylation-independent manner. The presence of NEDD8 is required to activate the CSN5 active site. Overall, by synergising cryo-EM with MS, we identified novel sensory regions of the CSN that mediate its stepwise activation mechanism and provide a framework for better understanding the regulatory mechanism of other Cullin family members.

2019-09-13 | PXD013001 | Pride

The dopamine transporter antiports potassium to increase the uptake of dopamine

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2022-05-20 | PXD032663 | Pride

Substrate-induced conformational dynamics of the dopamine transporter

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2019-11-12 | PXD013841 | Pride

Prion-like low complexity regions enable avid virus-host interactions during HIV-1 infection

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