Isolation and characterization of subunits of DB58, a lectin from the stems and leaves of Dolichos biflorus.
The DB58 lectin of the stems and leaves of Dolichos biflorus is a heterodimer composed of two closely related subunits, alpha and beta. These subunits were dissociated from one another in urea and isolated by high-performance anion-exchange chromatography. Steric exclusion chromatography of the isolated subunits in 6 M guanidine hydrochloride showed molecular weights of 30,900 and 29,800 for the alpha and beta subunits, respectively. The subunits have very similar amino acid compositions and are glycosylated at each of their two N-glycosylation consensus sites. Each of the subunits had weak carbohydrate binding activity. Reverse-phase chromatography of tryptic digests of the subunits showed identical peptide maps with the exception of peaks identified as COOH-terminal peptides. Analyses of these peptides, COOH-terminal amino acid analyses, and the small differences in amino acid composition between the 2 subunits establish that the beta subunit differs from the alpha subunit by the absence of 11 or 12 amino acids from its COOH terminus. This structural difference, combined with information from previous biosynthetic studies, establishes that the beta subunit is derived from the alpha subunit by posttranslational proteolytic modification at the COOH terminus. The heterogeneity in the extent of truncation suggests that this conversion occurs by sequential removal of amino acids rather than by endoproteolytic cleavage. The possible physiological significance of this modification is discussed.
Etzler
M E
ME
Section of Molecular and Cellular Biology, University of California, Davis 95616.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002846:Chromatography, Affinity; D007887:Fabaceae / Q000737:chemistry*; D037102:Lectins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D037121:Plant Lectins; D010946:Plants, Medicinal; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D000596:Amino Acids; C086819:DB58 lectin, Dolichos biflorus; D037102:Lectins; D037121:Plant Lectins