Diversity of odourant binding proteins revealed by an expressed sequence tag project on male Manduca sexta moth antennae

A small expressed sequence tag (EST) project generating 506 ESTs from 375 cDNAs was undertaken on the antennae of male Manduca sexta moths in an effort to discover olfactory receptor proteins. We encountered several clones that encode apparent transmembrane proteins; however, none is a clear candidate for an olfactory receptor. Instead we found a greater diversity of odourant binding proteins (OBPs) than previously known in moth antennae, raising the number known for M. sexta from three to seven. Together with evidence of seventeen members of the family from the Drosophila melanogaster genome project, our results suggest that insects may have many tens of OBPs expressed in subsets of the chemosensory sensilla on their antennae. These results support a model for insect olfaction in which OBPs selectively transport and present odourants to transmembrane olfactory receptors. We also found five members of a family of shorter proteins, named sensory appendage proteins (SAPs), that might also be involved in odourant transport. This small EST project also revealed several candidate odourant degrading enzymes including three P450 cytochromes, a glutathione S‐transferase and a uridine diphosphate (UDP) glucosyltransferase. Several first insect homologues of proteins known from vertebrates, the nematode Caenorhabditis elegans, yeast and bacteria were encountered, and most have now also been detected by the large D. melanogaster EST project. Only thirteen entirely novel proteins were encountered, some of which are likely to be cuticle proteins.

[1]  C DeLisi,et al.  The detection and classification of membrane-spanning proteins. , 1985, Biochimica et biophysica acta.

[2]  M. Capecchi,et al.  Targeted mutations in hoxa-9 and hoxb-9 reveal synergistic interactions. , 1997, Developmental biology.

[3]  K. Nakanishi,et al.  A photoaffinity-labelled insect sex pheromone for the moth Antheraea polyphemus , 1978, Nature.

[4]  M. Lerner,et al.  Molecular cloning and sequencing of general odorant-binding proteins GOBP1 and GOBP2 from the tobacco hawk moth Manduca sexta: comparisons with other insect OBPs and their signal peptides , 1991, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[5]  R. Vogt,et al.  Odorant-binding-protein subfamilies associate with distinct classes of olfactory receptor neurons in insects. , 1991, Journal of neurobiology.

[6]  C. Dulac,et al.  A Novel Family of Putative Pheromone Receptors in Mammals with a Topographically Organized and Sexually Dimorphic Distribution , 1997, Cell.

[7]  J. Carlson,et al.  Coexpression of Two Odorant-Binding Protein Homologs inDrosophila: Implications for Olfactory Coding , 1997, The Journal of Neuroscience.

[8]  G. Stange,et al.  Molecular cloning, by a novel approach, of a cDNA encoding a putative olfactory protein in the labial palps of the moth Cactoblastis cactorum. , 1997, Gene.

[9]  Dietmar Krautwurst,et al.  Identification of Ligands for Olfactory Receptors by Functional Expression of a Receptor Library , 1998, Cell.

[10]  J. Huet,et al.  Purification and characterization of multiple forms of odorant/pheromone binding proteins in the antennae of Mamestra brassicae (Noctuidae). , 1996, Insect biochemistry and molecular biology.

[11]  Cornelia I Bargmann,et al.  Reprogramming Chemotaxis Responses: Sensory Neurons Define Olfactory Preferences in C. elegans , 1997, Cell.

[12]  B. Hansson,et al.  Attracted or repelled?--a matter of two neurons, one pheromone binding protein, and a chiral center. , 1998, Biochemical and biophysical research communications.

[13]  H. Robertson Two large families of chemoreceptor genes in the nematodes Caenorhabditis elegans and Caenorhabditis briggsae reveal extensive gene duplication, diversification, movement, and intron loss. , 1998, Genome research.

[14]  H. Breer,et al.  Peripheral processes in insect olfaction. , 1992, Annual review of physiology.

[15]  J. Carlson,et al.  Olfaction in Drosophila: from odor to behavior. , 1996, Trends in genetics : TIG.

[16]  D. P. Smith,et al.  Identification of a Drosophila G protein alpha subunit (dGq alpha-3) expressed in chemosensory cells and central neurons. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[17]  R. Vogt,et al.  Intergeneric distribution and immunolocalization of a putative odorant-binding protein in true bugs (Hemiptera, Heteroptera). , 1998, The Journal of experimental biology.

[18]  K. Mikoshiba,et al.  Functional expression of a mammalian odorant receptor. , 1998, Science.

[19]  T. Kubo,et al.  Molecular cloning of cDNA for p10, a novel protein that increases in the regenerating legs of Periplaneta americana (American cockroach). , 1998, Insect biochemistry and molecular biology.

[20]  A. Taylor,et al.  A protein with several possible membrane-spanning domains encoded by the Drosophila segment polarity gene patched , 1989, Nature.

[21]  J. Lucchesi,et al.  Tosca: a Drosophila gene encoding a nuclease specifically expressed in the female germline. , 1996, Developmental biology.

[22]  Andrey Rzhetsky,et al.  A Spatial Map of Olfactory Receptor Expression in the Drosophila Antenna , 1999, Cell.

[23]  M. Rosbash,et al.  Members of a family of drosophila putative odorant-binding proteins are expressed in different subsets of olfactory hairs , 1994, Neuron.

[24]  S. Levy,et al.  The tetraspanin superfamily: molecular facilitators , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[25]  E. Mardis,et al.  Generation and analysis of 280,000 human expressed sequence tags. , 1996, Genome research.

[26]  John R. Carlson,et al.  A Novel Family of Divergent Seven-Transmembrane Proteins Candidate Odorant Receptors in Drosophila , 1999, Neuron.

[27]  C. Goodman,et al.  A Neural Tetraspanin, Encoded by late bloomer, That Facilitates Synapse Formation , 1996, Science.

[28]  M. Mavroidis,et al.  Cloning and characterization of a cDNA encoding a male‐specific serum protein of the Mediterranean fruit fly, Ceratitis capitata, with sequence similarity to odourant–binding proteins , 1998, Insect molecular biology.

[29]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[30]  Cori Bargmann,et al.  odr-10 Encodes a Seven Transmembrane Domain Olfactory Receptor Required for Responses to the Odorant Diacetyl , 1996, Cell.

[31]  Cori Bargmann,et al.  Divergent seven transmembrane receptors are candidate chemosensory receptors in C. elegans , 1995, Cell.

[32]  R A Steinbrecht,et al.  Are odorant-binding proteins involved in odorant discrimination? , 1996, Chemical senses.

[33]  N. Vickers,et al.  Combinatorial odor discrimination in the brain: Attractive and antagonist odor blends are represented in distinct combinations of uniquely identifiable glomeruli , 1998, The Journal of comparative neurology.

[34]  G. Hasan,et al.  Preferential Expression of Biotransformation Enzymes in the Olfactory Organs of Drosophila melanogaster, the Antennae* , 1999, The Journal of Biological Chemistry.

[35]  M. Ozaki,et al.  A putative lipophilic stimulant carrier protein commonly found in the taste and olfactory systems. A unique member of the pheromone-binding protein superfamily. , 1995, European journal of biochemistry.

[36]  D. P. Smith,et al.  LUSH odorant-binding protein mediates chemosensory responses to alcohols in Drosophila melanogaster. , 1998, Genetics.

[37]  H Breer,et al.  Cloning of genomic and complementary DNA encoding insect pheromone binding proteins: evidence for microdiversity. , 1991, Biochimica et biophysica acta.

[38]  Dean P. Smith,et al.  Olfactory mechanisms in Drosophila melanogaster , 1996, Current Opinion in Neurobiology.

[39]  R. Axel,et al.  A novel multigene family may encode odorant receptors: A molecular basis for odor recognition , 1991, Cell.

[40]  M. Maïbèche-Coisné,et al.  Pheromone binding proteins of the moth Mamestra brassicae: Specificity of ligand binding , 1997 .

[41]  H. Breer,et al.  Molecular cloning of an insect pheromone‐binding protein , 1989, FEBS letters.

[42]  H. Breer,et al.  Odorant binding proteins of Heliothis virescens. , 1993, Insect biochemistry and molecular biology.

[43]  L. Buck,et al.  A Multigene Family Encoding a Diverse Array of Putative Pheromone Receptors in Mammals , 1997, Cell.

[44]  L. Jennings,et al.  The activation of human platelets mediated by anti-human platelet p24/CD9 monoclonal antibodies. , 1990, The Journal of biological chemistry.

[45]  G. Prestwich,et al.  How is pheromone specificity encoded in proteins? , 1995, Chemical senses.

[46]  P. Pelosi,et al.  Binding proteins from the antennae of Bombyx mori. , 1996, Insect biochemistry and molecular biology.

[47]  Andrew Smith Genome sequence of the nematode C-elegans: A platform for investigating biology , 1998 .

[48]  C. Masson,et al.  Biochemical characterization, molecular cloning and localization of a putative odorant‐binding protein in the honey bee Apis mellifera L. (Hymenoptera: Apidea) , 1997, FEBS letters.

[49]  Glenn D. Prestwich,et al.  Patterns of Gene Duplication in Lepidopteran Pheromone Binding Proteins , 1998, Journal of Molecular Evolution.

[50]  M. Maïbèche-Coisné,et al.  Rapid Communication Molecular cloning of two pheromone binding proteins in the cabbage armyworm Mamestra brassicae , 1998 .

[51]  J. Sanes,et al.  Ontogeny of electroantennogram responses in the moth, Manduca sexta. , 1976, Journal of insect physiology.

[52]  P. Pelosi,et al.  Odorant-binding proteins in insects. , 1995, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[53]  J. Seehafer,et al.  Evidence that the signal-initiating membrane protein CD9 is associated with small GTP-binding proteins. , 1991, Biochemical and biophysical research communications.

[54]  R. Vogt,et al.  Snmp-1, a Novel Membrane Protein of Olfactory Neurons of the Silk Moth Antheraea polyphemus with Homology to the CD36 Family of Membrane Proteins* , 1997, The Journal of Biological Chemistry.

[55]  H. Breer,et al.  A novel class of binding proteins in the antennae of the silk moth Antheraea pernyi , 1990 .

[56]  M. Lerner,et al.  Characterization and cDNA cloning of the pheromone-binding protein from the tobacco hornworm, Manduca sexta: a tissue-specific developmentally regulated protein. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[57]  G. Paesen,et al.  The B proteins secreted by the tubular accessory sex glands of the male mealworm beetle, Tenebrio molitor, have sequence similarity to moth pheromone-binding proteins. , 1995, Insect biochemistry and molecular biology.

[58]  G. Prestwich,et al.  Expression and characterization of a lepidopteran general odorant binding protein. , 1997, Insect biochemistry and molecular biology.

[59]  P. Pelosi,et al.  Putative odorant-binding protein in antennae and legs of Carausius morosus (Insecta, Phasmatodea). , 1996, Insect biochemistry and molecular biology.

[60]  R. Axel,et al.  A novel family of genes encoding putative pheromone receptors in mammals , 1995, Cell.