Post-translational processing of two alpha-amylase inhibitors and an arcelin from the common bean, Phaseolus vulgaris.
Mass spectrometric methods were used to investigate the proteolytic processing and glycopeptide structures of three seed defensive proteins from Phaseolus vulgaris. The proteins were the alpha-amylase inhibitors alphaAI-1 and alphaAI-2 and arcelin-5, all of which are related to the seed lectins, PHA-E and PHA-L. The mass data showed that the proteolytic cleavage required for activation of the amylase inhibitors is followed by loss of the terminal Asn residue in alphaAI-1, and in all three proteins, seven or more residues were clipped from the C-termini, in the manner of the seed lectins. In most instances, individual glycoforms could be assigned at each Asn site, due to the unique masses of the plant glycopeptides. It was found that alphaAI-1 and alphaAI-2 differed significantly in their glycosylation patterns, despite their high sequence homology. These data complement the previous X-ray studies of the alpha1-amylase inhibitor and arcelin, where many of the C-terminal residues and glycopeptide residues could not be observed.
Young
N M
NM
Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ont. martin.young@nrc.ca
Thibault
P
P
Watson
D C
DC
Chrispeels
M J
MJ
D000595:Amino Acid Sequence; D004791:Enzyme Inhibitors; D007887:Fabaceae / Q000235:genetics / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins; D037102:Lectins / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D037121:Plant Lectins; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D010946:Plants, Medicinal; D011499:Protein Processing, Post-Translational; D000516:alpha-Amylases / Q000037:antagonists & inhibitors
D004791:Enzyme Inhibitors; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D037102:Lectins; D037121:Plant Lectins; D010940:Plant Proteins; C061510:alpha-amylase inhibitor, Phaseolus vulgaris; C090486:arcelin 5 protein, Phaesolus vulgaris; D000516:alpha-Amylases