1H, 13C, and 15N resonance assignment and secondary structure of the pheromone-binding protein2 from the agricultural pest Ostrinia furnacalis (OfurPBP2)

Dahal, Salik Ram; Lewellen, Jacob L.; Chaudhary, Bharat; Mohanty, Smita

doi:10.1007/s12104-020-09930-1

Cited by 2 publications

(9 citation statements)

References 20 publications

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“…Recombinant OfurPBP2 for the binding assay was expressed in bacteria and purified following the previously reported method. 14,17 Delipidation was carried out to remove the lipid bound to the protein from the bacterial cells. 21,24 The successful delipidation of OfurPBP2 was verified by 2D { 1 H, 15 N} HSQC (Figure S1).…”

Section: Pheromone-binding Affinitymentioning

confidence: 99%

“…Stereo views of the superposition of 20 lowest energy-minimized structures and a ribbon diagram of the overall structure are shown (Figure 4). The N-terminal segment (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22) forms an amphipathic helix (Figure 4, Figure S6A) but has slight distortion in the middle (at Tyr16) similar to AtraPBP1 23 and BmorPBP. 26 The most interesting and prominent feature in the OfurPBP2 structure is its C-terminus (131-143) with well-defined sequential d NN (i, i + 1) NOEs (Figure S5) forms an amphipathic helix (Figure S6B).…”

Section: Nmr Structure Determinationmentioning

confidence: 99%

“…All NMR data were acquired at 35 C on a Bruker Avance II 800 MHz spectrometer equipped with a triple resonance H/C/N cryoprobe TCI with pulse field gradients at the National High Magnetic Field Laboratory (NHMFL) at Tallahassee, FL. The following experiments were performed for sequential assignments 17 and structure analysis: 2D { 1 H, 15 N} HSQC, 2D { 1 H, 13 C} HSQC, 3D HNCA, 3D HN(CO)CA, 3D HNCO, 3D HN(CA)CO, 3D HNCACB, 3D CACB(CO)NH, 3D CC(CO)NH, 3D H(CCCO)NH, 3D HCCH-TOCSY, 3D 15 N-edited HSQC TOCSY, 3D 15 Nedited HSQC-NOESY (T m = 85 ms) and 3D 13 C-edited HSQC-NOESY (T m = 120 ms). In the 13 C-edited NOESY, the 13 C carrier frequency was set to 43 ppm for the aliphatic and 125 ppm for the aromatic region.…”

Section: Nmr Spectroscopy and Resonance Assignmentmentioning

confidence: 99%

“…Uniformly isotopically labeled ( 15 N and 15 N/ 13 C), recombinant OfurPBP2 was expressed in E.coli and purified by ion exchange and size exclusion chromatography as described previously. 14,17 NMR samples used for the structure determination contained 0.4 mM protein solution in 90% H 2 O/10% D 2 O in 50 mM phosphate buffer at pH 6.5 containing 1 mM EDTA and 0.1% NaN 3 .…”

Section: Sample Preparationmentioning

confidence: 99%

“…14,15,16 Despite high sequence similarity to other well-studied lepidopteran PBPs, 14,15,16 OfurPBP2 has remarkable differences in both biological gates (Figure 1). 8,17,14 Specifically, His70, which forms the histidine-gate with His95, is replaced by an arginine in OfurPBP2. Moreover, the C-terminal gate contains four additional charged residues, unlike other wellcharacterized lepidopteran PBPs 16,18,19 (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

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Ostrinia furnacalisPBP2 solution NMR structure: Insight into ligand binding and release mechanisms

et al. 2022

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Ostrinia furnacalis is an invasive lepidopteran agricultural pest that relies on olfaction for mating and reproduction. Male moths have an extremely sensitive olfactory system that can detect the sex pheromones emitted by females over a great distance. Pheromone‐binding proteins present in the male moth antenna play a key role in the pheromone uptake, transport, and release at the dendritic membrane of the olfactory neuron. Here, we report the first high‐resolution NMR structure of a pheromone‐binding protein from an Ostrinia species at pH 6.5. The core of the Ostrinia furnacalis PBP2 (OfurPBP2) consists of six helices, α1a (2–14), α1b (16–22), α2 (27–37), α3 (46–60), α4 (70–80), α5 (84–100), and α6 (107–124) surrounding a large hydrophobic pocket. The structure is stabilized by three disulfide bridges, 19–54, 50–108, and 97–117. In contrast to the unstructured C‐terminus of other lepidopteran PBPs, the C‐terminus of OfurPBP2 folds into an α‐helix (α7) at pH 6.5. The protein has nanomolar affinity towards both pheromone isomers. Molecular docking of both pheromones, E‐12 and Z‐12‐tetradecenyl acetate, to OfurPBP2 revealed that the residues Met5, Lys6, Met8, Thr9, Phe12, Phe36, Trp37, Phe76, Ser115, Phe118, Lys119, Ile122, His123, and Ala128 interact with both isomers, while Thr9 formed a hydrogen bond with the acetate head group. NMR structure and thermal unfolding studies with CD suggest that ligand release at pH 4.5 is likely due to the partial unfolding of the protein.

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Section: Pheromone-binding Affinitymentioning

confidence: 99%

Section: Nmr Structure Determinationmentioning

confidence: 99%

Section: Nmr Spectroscopy and Resonance Assignmentmentioning

confidence: 99%

Section: Sample Preparationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Ostrinia furnacalisPBP2 solution NMR structure: Insight into ligand binding and release mechanisms

et al. 2022

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NMR-Based Characterization of the Interaction between Yeast Oxa1-CTD and Ribosomes

Liu,

Yang,

Ruan

et al. 2023

IJMS

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In mitochondria, the major subunits of oxidative phosphorylation complexes are translated by the mitochondrial ribosome (mito-ribosome). The correct insertion and assembly of these subunits into the inner mitochondrial membrane (IMM) are facilitated by mitochondrial oxidase assembly protein 1 (Oxa1) during the translation process. This co-translational insertion process involves an association between the mito-ribosome and the C-terminus of Oxa1 (Oxa1-CTD) Nuclear magnetic resonance (NMR) methods were mainly used to investigate the structural characterization of yeast Oxa1-CTD and its mode of interaction with the E. coli 70S ribosome. Oxa1-CTD forms a transient α-helical structure within the residues P342–Q385, which were reported to form an α-helix when combining with the ribosome. Two conserved contact sites that could interact with the ribosome were further identified. The first site was located on the very end of the N-terminus (V321–I327), and the second one encompassed a stretch of amino acid residues I348–Q370. Based on our discoveries and previous reports, a model has been proposed in which Oxa1-CTD interacts with ribosomes, accompanied by transient-to-stable transitions at the second contact site. These observations may enhance our understanding of the potential role of Oxa1-CTD in facilitating the assembly of oxidative phosphorylation complexes and provide insight into the structural characteristics of Oxa1-CTD.

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1H, 13C, and 15N resonance assignment and secondary structure of the pheromone-binding protein2 from the agricultural pest Ostrinia furnacalis (OfurPBP2)

Cited by 2 publications

References 20 publications

Ostrinia furnacalisPBP2 solution NMR structure: Insight into ligand binding and release mechanisms

Ostrinia furnacalisPBP2 solution NMR structure: Insight into ligand binding and release mechanisms

NMR-Based Characterization of the Interaction between Yeast Oxa1-CTD and Ribosomes

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