Structural Basis of Ligand Binding and Release in Insect Pheromone-binding Proteins: NMR Structure of Antheraea polyphemus PBP1 at pH 4.5

• Published in: Journal of molecular biology 2007-11, Vol.373 (4), p.811-819
• Main Authors: Damberger, Fred F., Ishida, Yuko, Leal, Walter S., Wüthrich, Kurt
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antheraea polyphemus; Bombyx mori; Carrier Proteins - chemistry; Carrier Proteins - genetics; Hydrogen-Ion Concentration; Insect Proteins - chemistry; Insect Proteins - genetics; Magnetic Resonance Spectroscopy - methods; Models, Molecular; Molecular Sequence Data; Moths - genetics; Moths - metabolism; NMR structure; odorant-binding protein; olfaction; pH-dependent conformational transition; pheromone signaling; Protein Structure, Secondary; Sequence Homology, Amino Acid
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2007.07.078

The NMR structure of the Antheraea polyphemus pheromone-binding protein 1 at pH 4.5, ApolPBP1 A, was determined at 20 °C. The structure consists of six α-helices, which are arranged in a globular fold that encapsulates a central helix α7 formed by the C-terminal polypeptide segment 131–142. The 3D arrangement of these helices is anchored by the three disulfide bonds 19–54, 50–108 and 97–117, which were identified by NMR. Superposition of the ApolPBP1 A structure with the structure of the homologous pheromone-binding protein of Bombyx mori at pH 4.5, BmorPBP A, yielded an rmsd of 1.7 Å calculated for the backbone heavy-atoms N, C α and C′ of residues 10–142. In contrast, the present ApolPBP1 A structure is different from a recently proposed molecular model for a low-pH form of ApolPBP1 that does not contain the central helix α7. ApolPBP1 exhibits a pH-dependent transition between two different globular conformations in slow exchange on the NMR chemical shift timescale similar to BmorPBP, suggesting that the two proteins use the same mechanism of ligand binding and ejection. The extensive sequence homology observed for pheromone-binding proteins from moth species further implies that the previously proposed mechanism of ligand ejection involving the insertion of a C-terminal helix into the pheromone-binding site is a general feature of pheromone signaling in moths.