Accumulation of a lectin-like breakdown product of beta-conglutin catabolism in cotyledons of germinating Lupinus albus L. seeds.
During germination of Lupinus albus seeds, a 20-kDa polypeptide accumulates in the cotyledons of 4-d-old plants (Ferreira et al., 1995b, J Exp Bot 46: 211-219). Immunological, polypeptide cleavage with cyanogen bromide and amino acid sequencing experiments indicate that the 20-k-Da polypeptide and ubiquitin are structurally unrelated. However, there is a strong sequence homology between the 20-kDa polypeptide and the vicilin-like storage proteins from pea and soybean. Our results indicate that the 20-kDa polypeptide is an intermediate breakdown products of beta-conglutin catabolism, the vicilin-like storage protein from L. albus, and that its interaction with anti-ubiquitin antibodies results from the recognition of the antibodies by the 20-kDa polypeptide rather than by the opposite. Besides rabbit anti-ubiquitin antibodies, the 20-kDa polypeptide interacts with a variety of glycoproteins, including immunoglobulin G from several animal species, peroxidase and alkaline phosphatase, suggesting that it possess a lectin-type activity. Its activity is resistant to sodium dodecyl sulfate or methanol treatments, boiling and autoclaving. Purification of the 20-kDa polypeptide and immunological studies with anti-20-kDa-polypeptide antibodies showed that the non-glycosylated polypeptide is part of a glycoprotein with an estimated molecular mass of 210 kDa, composed of several types of structurally related subunit with molecular masses ranging from 14 to 50 kDa. Purified native protein containing the 20-kDa polypeptide self-aggregates in a calcium-dependent manner as reported for some glycosylated lectins. The possible physiological function of the 20-kDa polypeptide is discussed.
dos Ramos
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PC
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
Ferreira
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RM
Franco
E
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Teixeira
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AR
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002852:Chromatography, Ion Exchange; D018548:Cotyledon / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000502:physiology*; D015151:Immunoblotting; D037102:Lectins / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D037121:Plant Lectins; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010946:Plants, Medicinal; D011817:Rabbits; D051381:Rats; D055314:Seed Storage Proteins; D012639:Seeds / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000906:Antibodies; D037102:Lectins; D010446:Peptide Fragments; D037121:Plant Lectins; D010940:Plant Proteins; D055314:Seed Storage Proteins; C042457:conglutin protein, Lupinus angustifolius