Database of articles cited by EMDB/PDB/SASBDB data

Keywords
Structure methods	All X-ray diffraction NMR electron microscopy small angle scattering neutron diffraction fiber diffraction powder diffraction infrared spectroscopy EPR fluorescence transfer theoretical model Hybrid
Author
Journal
IF	>30 >20 >10 >5 all

Title	The carboxy terminus causes interfacial assembly of oleate hydratase on a membrane bilayer.
Journal, issue, pages	J Biol Chem, Vol. 300, Issue 2, Page 105627, Year 2024
Publish date	Jan 10, 2024
Authors	Christopher D Radka / Christy R Grace / Hale S Hasdemir / Yupeng Li / Carlos C Rodriguez / Patrick Rodrigues / Michael L Oldham / M Zuhaib Qayyum / Aaron Pitre / William J MacCain / Ravi C Kalathur / Emad Tajkhorshid / Charles O Rock /
PubMed Abstract	The soluble flavoprotein oleate hydratase (OhyA) hydrates the 9-cis double bond of unsaturated fatty acids. OhyA substrates are embedded in membrane bilayers; OhyA must remove the fatty acid from the ...The soluble flavoprotein oleate hydratase (OhyA) hydrates the 9-cis double bond of unsaturated fatty acids. OhyA substrates are embedded in membrane bilayers; OhyA must remove the fatty acid from the bilayer and enclose it in the active site. Here, we show that the positively charged helix-turn-helix motif in the carboxy terminus (CTD) is responsible for interacting with the negatively charged phosphatidylglycerol (PG) bilayer. Super-resolution microscopy of Staphylococcus aureus cells expressing green fluorescent protein fused to OhyA or the CTD sequence shows subcellular localization along the cellular boundary, indicating OhyA is membrane-associated and the CTD sequence is sufficient for membrane recruitment. Using cryo-electron microscopy, we solved the OhyA dimer structure and conducted 3D variability analysis of the reconstructions to assess CTD flexibility. Our surface plasmon resonance experiments corroborated that OhyA binds the PG bilayer with nanomolar affinity and we found the CTD sequence has intrinsic PG binding properties. We determined that the nuclear magnetic resonance structure of a peptide containing the CTD sequence resembles the OhyA crystal structure. We observed intermolecular NOE from PG liposome protons next to the phosphate group to the CTD peptide. The addition of paramagnetic MnCl indicated the CTD peptide binds the PG surface but does not insert into the bilayer. Molecular dynamics simulations, supported by site-directed mutagenesis experiments, identify key residues in the helix-turn-helix that drive membrane association. The data show that the OhyA CTD binds the phosphate layer of the PG surface to obtain bilayer-embedded unsaturated fatty acids.
External links	J Biol Chem / PubMed:38211817 / PubMed Central
Methods	EM (single particle) / NMR (solution)
Resolution	2.61 - 3.03 Å
Structure data	42480 8ur3 EMDB-42480, PDB-8ur3: Cryo-EM reconstruction of Staphylococcus aureus Oleate hydratase (OhyA) dimer with an ordered C-terminal membrane-association domain Method: EM (single particle) / Resolution: 2.61 Å 42484 8ur6 EMDB-42484, PDB-8ur6: Cryo-EM reconstruction of Staphylococcus aureus oleate hydratase (OhyA) dimer with a disordered C-terminal membrane-association domain Method: EM (single particle) / Resolution: 3.03 Å PDB-8um1: Structure of the Carboxy terminus of Oleate Hydratase Method: SOLUTION NMR PDB-8um2: Carboxy terminus of Oleate Hydratase in phosphate buffer Method: SOLUTION NMR
Source	staphylococcus aureus (bacteria)
Keywords	LIPID TRANSPORT / Oleate Hydratase / Membrane binding / HYDROLASE / oleate hydratase (OhyA) / phospholipids / membrane binding domain / amphipathic helices / interfacial enzyme / peripheral membrane protein