NMR structure of the unliganded <i>Bombyx mori</i> pheromone‐binding protein at physiological pH

Lee, Donghan; Damberger, Fred F.; Peng, Guihong; Horst, Reto; Güntert, Peter; Nikonova, Larisa; Leal, Walter S.; Wüthrich, Kurt

doi:10.1016/s0014-5793(02)03548-2

Cited by 91 publications

(128 citation statements)

References 34 publications

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“…6 Inset). Although not entirely surprising, given the size of this peptide (23), this observation indicates that the formation of the C-terminal ␣-helix might be stabilized by interactions of the terminal residues with residues forming the binding cavity (19,22). Except for three acidic residues, the terminal fragment is composed almost entirely of hydrophobic amino acid residues.…”

Section: Resultsmentioning

confidence: 87%

“…Previously, we suggested that the loss of binding affinity at low pH is due to the occupation of the binding cavity by a C-terminal ␣-helix in Bmor-PBP A (Fig. 3A) (18,19). To test this hypothesis, we expressed a truncated form of BmorPBP B by removing the segment P129-SMDVAVGEILAE-V142 (see Fig.…”

Section: Resultsmentioning

confidence: 97%

“…5, which is published as supporting information on the PNAS web site), which is involved in the formation of the C-terminal ␣-helix (MDVAVGEILAE) (18). The molecular mass of BmorPBP⌬P129-V142 obtained by LC-ESI͞MS (observed, 14,466 Da; calculated, 14,472 Da) indicated that all Cys residues were linked to form three disulfide bridges, as in the native conformation (18)(19)(20)(21)(22). CD data (see Fig.…”

Section: Resultsmentioning

confidence: 98%

“…This corresponds to a t 1/2 of the ligand-protein complex of Ϸ100 s, an extremely slow process given the dynamics of the insect olfactory system. However, it has been strongly suggested that the release of pheromone is triggered by a pH-dependent conformational change (8,18,19,22,29), and this is a very fast process (t 1/2 Ϸ 9 ms). The data presented here thus support the hypothesis that PBPs are essential for the dynamics of the insect olfactory system.…”

Section: Discussionmentioning

confidence: 99%

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Kinetics and molecular properties of pheromone binding and release

Leal

Chen

Ishida

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

164

139

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Transient kinetic studies have shown that the uptake of the pheromone (bombykol) of the silkworm moth (Bombyx mori), by its pheromone-binding protein (PBP) BmorPBP, proceeds with an ''on'' rate of 0.068 ؎ 0.01 M ؊1 ⅐s ؊1 . With the high concentration of PBP in the sensillar lymph (10 mM), the half-life for the uptake of pheromone in vivo is Ϸ1 ms. A pH-dependent conformational change (BmorPBP B 3 BmorPBP A ), associated with the release of pheromone, is a first-order reaction (k ‫؍‬ 74.1 ؎ 0.32 s ؊1 ; t1/2, 9.3 ms). Under physiological conditions, both reactions proceed with half-life times on the order of milliseconds, as is required for odorant-oriented navigation in insects. Molecular interactions of bombykol with both native and mutated PBPs were analyzed by a novel binding assay. A recombinant protein with the native conformation (BmorPBP) showed high binding affinity (K D ‫؍‬ 105 nM) at pH 7 but low affinity (K D ‫؍‬ 1,600 nM) at pH 5, when tested at both low and high KCl concentrations. A protein with a C-terminal segment deleted (BmorPBP⌬P129-V142) was found to bind bombykol at pH 7 and at pH 5 with the same affinity as the native protein at pH 7, indicating that the C-terminal segment is essential for preventing binding at low pH. Binding studies with three mutated proteins (BmorPBPW37F, BmorPBPW127F, and BmorPBPW37A) showed that replacing Trp-37 (with Phe or Ala) or Trp-127 (with Phe) did not affect the binding affinity to bombykol. Fluorescence studies shed light on the contributions of Trp-37 and Trp-127 emissions to the overall fluorescence.Bombyx mori pheromone-binding protein ͉ bombykol ͉ effect of C terminus on pheromone release ͉ fast uptake of pheromone and delivery ͉ mutated pheromone-binding proteins F or many insects, small-molecule signals communicate the availability of food, the presence of friends and foes, and the readiness to mate. In general, mate-finding is an essential prerequisite for exploring insects' enormous reproductive potential and, consequently, leads to their domination of the terrestrial world. To advertise their readiness to mate, female moths, for example, produce and release sex pheromones. Only minute amounts are released, so as to avoid chemical conspicuousness. Once released, the chemical signals are diluted in the environment and mixed with a myriad of physiologically irrelevant compounds, including pheromones from other species. Even though each species communicates with a specific pheromonic language, males can detect the low-level signals from conspecific females because of a highly developed olfactory system. To find females and successfully reproduce, males may have to make long-distance, odorant-oriented flights. Navigation requires a highly selective, sensitive, and dynamic sensory system for the detection of pheromones. Given the structure of pheromone plumes (1), insects have only a few milliseconds to reset their detectors while flying in the clean air spaces between pockets of chemical signals.Pheromones are largely hydrophobic compounds, whereas pheromo...

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Section: Resultsmentioning

confidence: 87%

Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

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Kinetics and molecular properties of pheromone binding and release

Leal

Chen

Ishida

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

164

139

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“…Conformational heterogeneity has been observed in several other OBPs (14,17,59,60) and is most often associated with residues that regulate access to the ligand binding pocket. However, this heterogeneity is not nearly as extensive as that seen with OBP4 and LUSH.…”

Section: Discussionmentioning

confidence: 99%

New Insights into the Mechanism of Odorant Detection by the Malaria-transmitting Mosquito Anopheles gambiae

Davrazou

Dong

Murphy

et al. 2011

Journal of Biological Chemistry

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Anopheles gambiae mosquitoes that transmit Plasmodium falciparum malaria use a series of olfactory cues present in human sweat to locate their hosts for a blood meal. Recognition of these odor cues occurs through the interplay of odorant receptors and odorant-binding proteins (OBPs) that bind to odorant molecules and transport and present them to the receptors. Recent studies have implicated potential heterodimeric interactions between two OBPs, OBP1 and OBP4, as important for perception of indole by the mosquito (Biessmann, H., Andronopoulou, E., Biessmann, M. R., Douris, V., Dimitratos, S. D., Eliopoulos, E., Guerin, P. M., Iatrou, K., Justice, R. W., Kröber, T., Marinotti, O., Tsitoura, P., Woods, D. F., and Walter, M. F. (2010) PLoS ONE 5, e9471; Qiao, H., He, X., Schymura, D., Ban, L., Field, L., Dani, F. R., Michelucci, E., Caputo, B., della Torre, A., Iatrou, K., Zhou, J. J., Krieger, J., and Pelosi, P. (2011) Cell. Mol. Life Sci. 68, 1799 -1813). Here we present the 2.0 Å crystal structure of the OBP4-indole complex, which adopts a classical odorant-binding protein fold, with indole bound at one end of a central hydrophobic cavity. Solution-based NMR studies reveal that OBP4 exists in a molten globule state and binding of indole induces a dramatic conformational shift to a well ordered structure, and this leads to the formation of the binding site for OBP1. Analysis of the OBP4-OBP1 interaction reveals a network of contacts between residues in the OBP1 binding site and the core of the protein and suggests how the interaction of the two proteins can alter the binding affinity for ligands. These studies provide evidence that conformational ordering plays a key role in regulating heteromeric interactions between OBPs.Anopheles gambiae mosquitoes are the primary vectors for malaria caused by Plasmodium falciparum and have an extremely high preference for feeding on human hosts (1, 2) and are attracted to odor molecules from incubated human sweat and other skin emanations (3-6). Disrupting the normal olfactory responses to these odors presents an attractive tool to combat transmission of malaria and other mosquito borne diseases, and a number of efforts are now under way to discover novel reagents for this purpose.In insects the detection of odorants occurs primarily in the olfactory sensilla and involves the interplay of membranebound olfactory receptors (7) and odorant-binding proteins (OBPs), 3 which are expressed into the lymph fluid that surrounds the olfactory dendrites (8) where they can reach concentrations in the millimolar range (9, 10). OBPs have multiple roles including protecting odors from degradation and transporting them to the olfactory receptors (11, 12). There is evidence that OBPs have two primary roles in odorant perception. In the first model a number of groups have proposed that OBPs act as passive carriers for the odorant, and pH changes in the vicinity of the dendritic membrane lead to conformational changes that stimulate ligand release, freeing the ligand to activate the receptor (1...

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Structural Recognition Between Odorants, Olfactory‐Binding Proteins and Olfactory Receptors ‐ First Events in Odour Coding

Pernollet¹,

Briand²

2004

Flavor Perception

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NMR structure of the unliganded Bombyx mori pheromone‐binding protein at physiological pH

Cited by 91 publications

References 34 publications

Kinetics and molecular properties of pheromone binding and release

Kinetics and molecular properties of pheromone binding and release

New Insights into the Mechanism of Odorant Detection by the Malaria-transmitting Mosquito Anopheles gambiae

Structural Recognition Between Odorants, Olfactory‐Binding Proteins and Olfactory Receptors ‐ First Events in Odour Coding

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