Purification, structural characterization, cloning and immunocytochemical localization of chemoreception proteins from Schistocerca gregaria

被引:243
作者
Angeli, S
Ceron, F
Scaloni, A
Monti, M
Monteforti, G
Minnocci, A
Petacchi, R
Pelosi, P
机构
[1] Univ Pisa, Dipartimento Chim & Biotecnol Agrarie, I-56124 Pisa, Italy
[2] Univ Pisa, Scuola Super Studi Perfezionamento S Anna, Pisa, Italy
[3] Natl Res Council, Ctr Int Serv Spettrometria Massa IABBAM, Naples, Italy
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1999年 / 262卷 / 03期
关键词
chemosensory proteins; contact sensilla; disulfide bridges; Schistocerca gregaria; sequence analysis;
D O I
10.1046/j.1432-1327.1999.00438.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 end sequenced. Their expression products are polypeptide chains of 109 amino acids shelling 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 a ere 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.
引用
收藏
页码:745 / 754
页数:10
相关论文
共 55 条
[1]  
BLANEY WM, 1969, J ZOOL, V157, P509
[2]   A NOVEL CLASS OF BINDING-PROTEINS IN THE ANTENNAE OF THE SILK MOTH ANTHERAEA-PERNYI [J].
BREER, H ;
KRIEGER, J ;
RAMING, K .
INSECT BIOCHEMISTRY, 1990, 20 (07) :735-740
[3]   LIGAND-BINDING CHARACTERISTICS OF HOMOLOGOUS RAT AND MOUSE URINARY PROTEINS AND PYRAZINE-BINDING PROTEIN OF CALF [J].
CAVAGGIONI, A ;
FINDLAY, JBC ;
TIRINDELLI, R .
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, 1990, 96 (03) :513-520
[4]  
Chapman R.F., 1990, BIOL GRASSHOPPERS
[6]   Separation, characterization and sexual heterogeneity of multiple putative odorant-binding proteins in the honeybee Apis mellifera L. (Hymenoptera: Apidea) [J].
Danty, E ;
Arnold, G ;
Huet, JC ;
Huet, D ;
Masson, C ;
Pernollet, JC .
CHEMICAL SENSES, 1998, 23 (01) :83-91
[7]   OLFACTION IN A HEMIMETABOLOUS INSECT - ANTENNAL-SPECIFIC PROTEIN IN ADULT LYGUS-LINEOLARIS (HETEROPTERA, MIRIDAE) [J].
DICKENS, JC ;
CALLAHAN, FE ;
WERGIN, WP ;
ERBE, EF .
JOURNAL OF INSECT PHYSIOLOGY, 1995, 41 (10) :857-867
[8]  
DINH BAO-LMH, 1965, J IMMUNOL, V95, P574
[9]   ODORANT BINDING BY A PHEROMONE BINDING-PROTEIN - ACTIVE-SITE MAPPING BY PHOTOAFFINITY-LABELING [J].
DU, GH ;
NG, CS ;
PRESTWICH, GD .
BIOCHEMISTRY, 1994, 33 (16) :4812-4819
[10]   MAJOR URINARY PROTEIN COMPLEX OF NORMAL MICE - ORIGIN [J].
FINLAYSON, JS ;
ASOFSKY, R ;
POTTER, M ;
RUNNER, CC .
SCIENCE, 1965, 149 (3687) :981-+