Chemical Senses Advance Access published online on November 16, 2009
Chemical Senses, doi:10.1093/chemse/bjp073
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The Soluble Proteome of the Drosophila Antenna
1 Departments of Biology and Genetics and W. M. Keck Center for Behavioral Biology, Box 7617, North Carolina State University, Raleigh, NC 27695-7617, USA 2 Department of Chemistry, Box 8204, North Carolina State University, Raleigh, NC 27695-8204, USA
Correspondence to be sent to: Robert R.H. Anholt, Department of Biology, Box 7617, North Carolina State University, Raleigh, NC 27695-7617, USA. e-mail: anholt{at}ncsu.edu
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Abstract |
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The olfactory system of Drosophila melanogaster is one of the best characterized chemosensory systems. Identification of proteins contained in the third antennal segment, the main olfactory organ, has previously relied primarily on immunohistochemistry, and although such studies and in situ hybridization studies are informative, they focus generally on one or few gene products at a time, and quantification is difficult. In addition, purification of native proteins from the antenna is challenging because it is small and encased in a hard cuticle. Here, we describe a simple method for the large-scale detection of soluble proteins from the Drosophila antenna by chromatographic separation of tryptic peptides followed by tandem mass spectrometry with femtomole detection sensitivities. Examination of the identities of these proteins indicates that they originate both from the extracellular perilymph and from the cytoplasm of disrupted cells. We identified enzymes involved with intermediary metabolism, proteins associated with regulation of gene expression, nucleic acid metabolism and protein metabolism, proteins associated with microtubular transport, 8 odorant-binding proteins, protective enzymes associated with antibacterial defense and defense against oxidative damage, cuticular proteins, and proteins of unknown function, which represented about one-third of all soluble proteins. The procedure described here opens the way for precise quantification of any target protein in the Drosophila antenna and should be readily applicable to antennae from other insects.
Key words: chemosensation, mass spectrometry, odorant-binding proteins, olfaction, proteomics
Accepted 21 September 2009