The complete sequence of the acidic subunit from Mojave toxin determined by Edman degradation and mass spectrometry.
Mojave toxin, a heterodimeric, neurotoxic phospholipase complex from Crotalus scutulatus scutulatus, is one of a group of closely related rattlesnake toxins for which much structural information is still lacking. The complete amino-acid sequence of the acidic subunit from Mojave toxin was determined. The three individual peptide chains, derived from the acidic subunit by reductive alkylation, were separated by high-performance liquid chromatography. Fragmentations of the A and B chains were done using specific proteinases and the resulting peptide mixtures were fractionated by reverse-phase high-performance liquid chromatography. Sequence analyses on the intact chains and the fragments from digests were done by automated Edman degradation, carboxypeptidase Y degradation and triple-quadrupole and tandem-quadrupole Fourier-transform mass spectrometry. The sequence for each acidic subunit chain is very similar to the corresponding chain from the related neurotoxin complex, crotoxin, and overall the sequence is similar to the sequences of group I and II phospholipases A2. The N-terminus of the B chain is blocked by pyroglutamic acid. The existence of two distinct and closely related C chains was established. It is unlikely that the small sequence difference can account for the isoforms that are present in purified Mojave toxin and in unfractionated venom.
Bieber
A L
AL
Department of Chemistry, Arizona State University, Tempe 85287.
Becker
R R
RR
McParland
R
R
Hunt
D F
DF
Shabanowitz
J
J
Yates
J R
JR
Martino
P A
PA
Johnson
G R
GR
D000595:Amino Acid Sequence; D002268:Carboxypeptidases; D002851:Chromatography, High Pressure Liquid; D003435:Crotalid Venoms; D003439:Crotoxin; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009498:Neurotoxins; D010446:Peptide Fragments; D010741:Phospholipases A
D003435:Crotalid Venoms; C018107:Mojave toxin; D009498:Neurotoxins; D010446:Peptide Fragments; D003439:Crotoxin; D010741:Phospholipases A; D002268:Carboxypeptidases; C022633:serine carboxypeptidase