Complete amino acid sequence of the Aa6 subunit of the scorpion Androctonus australis hemocyanin determined by Edman degradation and mass spectrometry.
The primary structure of the hemocyanin 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 (71,890 +/- 7 Da) is about 30 Da higher than the mass calculated from the sequence data (71,860.1 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 alpha 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.
Buzy
A
A
Institut de Biologie Structurale Jean-Pierre Ebel (CEA-CNRS), Grenoble, France.
Gagnon
J
J
Lamy
J
J
Thibault
P
P
Forest
E
E
Hudry-Clergeon
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001181:Arthropods / Q000235:genetics; D003488:Cyanogen Bromide; D006433:Hemocyanins / Q000737:chemistry* / Q000235:genetics*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011487:Protein Conformation; D012605:Scorpions / Q000737:chemistry* / Q000235:genetics*; D017386:Sequence Homology, Amino Acid
D010446:Peptide Fragments; D006433:Hemocyanins; D003488:Cyanogen Bromide