COMPLETE AMINO-ACID-SEQUENCE OF THE AA6 SUBUNIT OF THE SCORPION ANDROCTONUS-AUSTRALIS HEMOCYANIN DETERMINED BY EDMAN DEGRADATION AND MASS-SPECTROMETRY

被引:25
作者
BUZY, A
GAGNON, J
LAMY, J
THIBAULT, P
FOREST, E
HUDRYCLERGEON, G
机构
[1] CEA, CNRS, INST BIOL STRUCT JEAN PIERRE EBEL, F-38027 GRENOBLE, FRANCE
[2] FAC PHARM TOURS, BIOCHIM FONDAMENTALE LAB, TOURS, FRANCE
[3] CNRS, URA 1334, TOURS, FRANCE
[4] NATL RES COUNCIL CANADA, INST MARINE BIOSCI, HALIFAX, NS, CANADA
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1995年 / 233卷 / 01期
关键词
HEMOCYANIN; ANDROCTONUS AUSTRALIS; CHEMICAL SEQUENCING; MASS SPECTROMETRY;
D O I
10.1111/j.1432-1033.1995.093_1.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
The primary structure of the hernocyanin Aa6 subunit from the scorpion Androctonus australis was resolved by using protein sequencing and mass spectrometry for analysis of the polypeptide chain and of fragments obtained by CNBr, trypsin, and chymotrypsin cleavage. Due to the high sensitivity of the methodologies used, only a small amount of material, less than 1 mg, was consumed. The complete sequence is composed of 626 amino acid residues and the protein is not glycosylated but probably phosphorylated at Ser374. Its molecular mass measured by mass spectrometry (71890 +/- 7 Da) is about 30 Da higher than the mass calculated from the sequence data (71860.1 Da). The origin of this, difference is not clear but could result from minor molecular heterogeneities. Within the chelicerates, the Aa6 subunit of the arachnid A. australis shares 405 identical residues with chain e of another arachnid, Eurypelma californicum, and 399 with chain a of the merostom Tachypleus tridentatus. The degrees of identity between these three subunits, which are known to occupy the same location in the native hemocyanin oligomers, are significantly higher than those existing between the subunits a, d, and e of E. californicum. This favors the hypothesis that gene duplications, leading to separate chains in one species, have occurred before the divergence between arachnids and merostoms.
引用
收藏
页码:93 / 101
页数:9
相关论文
共 32 条
[1]   PANULIRUS-INTERRUPTUS HEMOCYANIN - THE ELUCIDATION OF THE COMPLETE AMINO-ACID-SEQUENCE OF SUBUNIT-A [J].
BAK, HJ ;
BEINTEMA, JJ .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1987, 169 (02) :333-348
[2]  
BEINTEMA JJ, 1994, MOL BIOL EVOL, V11, P493
[3]   MASS-SPECTROMETRY [J].
BURLINGAME, AL ;
BAILLIE, TA ;
RUSSELL, DH .
ANALYTICAL CHEMISTRY, 1992, 64 (12) :R467-R502
[4]   INTEGRATION OF MASS-SPECTROMETRY IN ANALYTICAL BIOTECHNOLOGY [J].
CARR, SA ;
HEMLING, ME ;
BEAN, MF ;
ROBERTS, GD .
ANALYTICAL CHEMISTRY, 1991, 63 (24) :2802-2824
[5]   PRIMARY STRUCTURE OF THE OLIGOSACCHARIDE MOIETY OF HEMOCYANIN FROM THE SCORPION ANDROCTONUS-AUSTRALIS [J].
DEBEIRE, P ;
MONTREUIL, J ;
GOYFFON, M ;
VANKUIK, AJ ;
VANHALBEEK, H ;
VLIEGENTHART, JFG .
CARBOHYDRATE RESEARCH, 1986, 151 :305-310
[6]   ELECTROSPRAY IONIZATION FOR MASS-SPECTROMETRY OF LARGE BIOMOLECULES [J].
FENN, JB ;
MANN, M ;
MENG, CK ;
WONG, SF ;
WHITEHOUSE, CM .
SCIENCE, 1989, 246 (4926) :64-71
[7]   3.2 A STRUCTURE OF THE COPPER-CONTAINING, OXYGEN-CARRYING PROTEIN PANULIRUS-INTERRUPTUS HEMOCYANIN [J].
GAYKEMA, WPJ ;
HOL, WGJ ;
VEREIJKEN, JM ;
SOETER, NM ;
BAK, HJ ;
BEINTEMA, JJ .
NATURE, 1984, 309 (5963) :23-29
[8]  
HAZES B, 1993, PROTEIN SCI, V2, P597
[9]   HIGHER-ORDER ASSEMBLIES OF MOLLUSCAN HEMOCYANINS [J].
HERSKOVITS, TT ;
HAMILTON, MG .
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, 1991, 99 (01) :19-34
[10]   MOLECULAR-WEIGHT ANALYSIS OF PROTEINS [J].
JARDINE, I .
METHODS IN ENZYMOLOGY, 1990, 193 :441-455