The carboxy terminus causes interfacial assembly of oleate hydratase on a membrane bilayer

• DOI: 10.1016/j.jbc.2024.105627
• Publication Date: 2024-01-10T02:05:56Z
• 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
• Keywords: Biotransformation; Pseudomonas syringae

The soluble flavoprotein oleate hydratase (OhyA) hydrates the 9-cis double bond of unsaturated fatty acids. OhyA substrates are embedded in membrane bilayers; must remove acid from bilayer and enclose it active site. Here, we show that positively charged helix-turn-helix motif carboxy terminus (CTD) is responsible for interacting with negatively phosphatidylglycerol (PG) bilayer. Super-resolution microscopy Staphylococcus aureus cells expressing green fluorescent protein fused to or CTD sequence shows subcellular localization along cellular boundary, indicating membrane-associated sufficient recruitment. Using cryo-electron microscopy, solved dimer structure conducted 3D variability analysis reconstructions assess flexibility. Our surface plasmon resonance experiments corroborated binds PG nanomolar affinity found has intrinsic binding properties. We determined nuclear magnetic a peptide containing resembles crystal structure. observed intermolecular NOE liposome protons next phosphate group peptide. addition paramagnetic MnCl