Purification, structural characterization, cloning and immunocytochemical localization of chemoreception proteins from Schistocerca gregaria.
Soluble low-molecular-mass protein isoforms were purified from chemosensory organs (antennae, tarsi and labrum) of the desert locust Schistocerca gregaria. Five genes encoding proteins of this group were amplified by PCR from cDNAs of tarsi and sequenced. Their expression products are polypeptide chains of 109 amino acids showing 40-50% sequence identity with putative olfactory proteins from Drosophila melanogaster and Cactoblastis cactorum. Direct structural investigation on isoforms purified from chemosensory organs revealed the presence in the expression products of two of the genes cloned. Two additional protein isoforms were detected and their molecular structure exhaustively characterized. MS analysis of all isoforms demonstrated that the four cysteine residues conserved in the polypeptide chain were involved in disulfide bridges (Cys29-Cys38 and Cys57-Cys60) and indicated the absence of any additional post-translational modifications. Immunocytochemistry experiments, performed with rabbit antiserum raised against the protein isoform mixture, showed selective labelling of the outer lymph in contact sensilla of tarsi, maxillary palps and antennae. Other types of sensilla were not labelled, nor were the cuticle and dendrites of the sensory cells. No binding of radioactively labelled glucose or bicarbonate was detected, in disagreement with the hypothesis that this class of proteins is involved in the CO2-sensing cascade. Our experimental data suggest that the proteins described here could be involved in contact chemoreception in Orthoptera.
Angeli
S
S
Scuola Superiore di Studi Universitari e di Perfezionamento S. Anna, Pisa, Italy.
Ceron
F
F
Scaloni
A
A
Monti
M
M
Monteforti
G
G
Minnocci
A
A
Petacchi
R
R
Pelosi
P
P
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002628:Chemoreceptor Cells / Q000737:chemistry*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D006110:Grasshoppers / Q000737:chemistry* / Q000648:ultrastructure; D007106:Immune Sera / Q000096:biosynthesis; D007150:Immunohistochemistry; D019476:Insect Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018076:DNA, Complementary; D007106:Immune Sera; D019476:Insect Proteins; D020033:Protein Isoforms