Assignment of the five disulfide bridges in an alpha-amylase inhibitor from wheat kernel by fast-atom-bombardment mass spectrometry and Edman degradation.
The assignment of the five disulfide bridges in an alpha-amylase monomeric inhibitor from wheat kernel (coded 0.28) was achieved by combining fast-atom-bombardment mass spectrometry (FAB-MS) and automatic sequencing based on Edman degradation. Direct FAB-MS analysis of the native and reduced enzymatic digests of the protein allowed the assignment of three disulfide bridges out of five, including those involving two adjacent cysteine residues. The remaining two disulfide bridges were assigned by sequencing automatically the peptide clusters purified from the tryptic digest of the native protein.
Poerio
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E
Dipartimento di Agrobiologia e Agrochimica, Università della Tuscia, Viterbo, Italy.
Caporale
C
C
Carrano
L
L
Pucci
P
P
Buonocore
V
V
D000595:Amino Acid Sequence; D004220:Disulfides / Q000032:analysis; D010450:Endopeptidases; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D010449:Peptide Mapping; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D011487:Protein Conformation; D016339:Spectrometry, Mass, Fast Atom Bombardment; D014908:Triticum; D014357:Trypsin; D000516:alpha-Amylases / Q000037:antagonists & inhibitors*
D004220:Disulfides; D010446:Peptide Fragments; D010940:Plant Proteins; C060586:WDAI-3 protein, Triticum aestivum; D000516:alpha-Amylases; D010450:Endopeptidases; D014357:Trypsin