Characterization of a subfamily of beetle odorant-binding proteins found in hemolymph.
In insects, hydrophobic odorants are transported through the sensillar lymph to receptors on sensory neurons by odorant-binding proteins (OBPs). The beetle Tenebrio molitor, which is a pest of stored grain products, produces a set of 12-14-kDa OBP-like proteins in its hemolymph. The structure of one of these proteins and that of a moth pheromone-binding protein have been solved. Both proteins have at least six alpha-helices with an internal, hydrophobic, ligand-binding pocket, but the beetle OBP lacks one of the disulfide bonds immediately adjacent to this pocket. To explore this difference and to sample isoform diversity, T. molitor hemolymph OBPs were fractionated by size-exclusion chromatography and reversed-phase high performance liquid chromatography. Selected fractions were reduced and alkylated, and tryptic peptides were sequenced by tandem mass spectrometry. Partial sequences of 7 different isoforms were obtained and used to clone 9 new cDNAs encoding OBPs with identities from 32 to 99%. The more divergent isoforms have numerous substitutions of hydrophobic residues that presumably alter the shape and specificity of the ligand-binding pocket. These isoforms all lack the same third disulfide bridge and are more similar to one another than to any of the 38 OBPs in Drosophila melanogaster. They have presumably arisen via gene duplication following separation of the major insect orders.
Graham
Laurie A
LA
Department of Biochemistry, Queen's University, Kingston, Ontario K7L 3N6, Canada. grahamla@post.queensu.ca
Brewer
Dyanne
D
Lajoie
Gilles
G
Davies
Peter L
PL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006458:Hemolymph / Q000378:metabolism; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009812:Odorants; D010802:Phylogeny; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D018035:Receptors, Odorant / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D013712:Tenebrio / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D019476:Insect Proteins; D020033:Protein Isoforms; D018035:Receptors, Odorant; C045648:odorant-binding protein