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Ligand binding to six recombinant pheromone-binding proteins of Antheraea polyphemus and Antheraea pernyi

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Abstract

Binding properties of six heterologously expressed pheromone-binding proteins (PBPs) identified in the silkmoths Antheraea polyphemus and Antheraea pernyi were studied using tritium-labelled pheromone components, (E,Z)-6,11-hexadecadienyl acetate (3H-Ac1) and (E,Z)-6,11-hexadecadienal (3H-Ald), common to both species. In addition, a known ligand of PBP and inhibitor of pheromone receptor cells, the tritium-labelled esterase inhibitor decyl-thio-1,1,1-trifluoropropanone (3H-DTFP), was tested. The binding of ligands was measured after native gel electrophoresis and cutting gel slices. In both species, PBP1 and PBP3 showed binding of 3H-Ac1. In competition experiments with 3H-Ac1 and the third unlabelled pheromone component, (E,Z)-4,9-tetradecadienyl acetate (Ac2), the PBP1 showed preferential binding of Ac1, whereas PBP3 preferentially bound Ac2. The PBP2 of both species bound 3H-Ald only. All of the six PBPs strongly bound 3H-DTFP. Among unlabelled pheromone derivatives, alcohols were revealed to be the best competitors for 3H-Ac1 and 3H-Ald bound to PBPs. No pH influence was found for 3H-Ac1 binding to, or its release from, the PBP3 of A. polyphemus and A. pernyi between pH 4.0 and pH 7.5. The data indicate binding preference of each of the three PBP-subtypes (1–3) for a specific pheromone component and support the idea that PBPs contribute to odour discrimination, although to a smaller extent than receptor activation.

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Abbreviations

Ac1 :

(E,Z)-6,11-hexadecadienyl acetate

Ac2 :

(E,Z)-4,9-tetradecadienyl acetate

Ald :

(E,Z)-6,11-hexadecadienal

AMA :

1-amino-anthracene

cpm :

counts per min

DTFP :

decyl-thio-1,1,1-trifluoropropanone

ES-MS :

electrospray mass spectrometry

OH:

(E,Z)-6,11-hexadecadienol

PAGE :

polyacrylamide gel electrophoresis

PCR :

polymerase chain reaction

PBP :

pheromone-binding protein

SDS :

sodium dodecyl sulphate

Z-11 OH :

Z-11 hexadecenol

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Acknowledgements

We are indebted to Dr. J. Krieger for providing the recombinant PBPs of A. polyphemus and A. pernyi and the delipidated PBP1 of A. polyphemus, and for discussion. We thank R.A. Steinbrecht and B. Pophof for comments and A. Biederman-Thorson for linguistic help.

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  1. Max-Planck-Institut fuer Verhaltensphysiologie Seewiesen, 82319, Starnberg, Germany

    R. Maida, G. Ziegelberger & K.-E. Kaissling

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  1. R. Maida
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  2. G. Ziegelberger
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  3. K.-E. Kaissling
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Correspondence to K.-E. Kaissling.

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Communicated by G. Heldmaier

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Maida, R., Ziegelberger, G. & Kaissling, KE. Ligand binding to six recombinant pheromone-binding proteins of Antheraea polyphemus and Antheraea pernyi . J Comp Physiol B 173, 565–573 (2003). https://doi.org/10.1007/s00360-003-0366-4

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