Journal of Comparative Physiology B

, Volume 173, Issue 7, pp 565–573 | Cite as

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

Original Paper

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.

Keywords

Insect olfaction Pheromone-binding proteins Insect pheromones pH dependence of pheromone binding Decyl-thio-1,1,1-trifluoropropanone 

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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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Max-Planck-Institut fuer Verhaltensphysiologie SeewiesenStarnbergGermany

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