Characterization of the endoplasmic reticulum-associated precursor of concanavalin A. Partial amino acid sequence and lectin activity.
Concanavalin A (ConA), which is not a glycoprotein, is synthesized as a glycoprotein precursor (pro-ConA) which is post-translationally processed. This processing results in the loss of a small glycopeptide with a high mannose oligosaccharide. Carrington et al. (Carrington, D.M., Auffret, A., and Hanke, D.E. (1985) Nature 313, 64-66) determined the nucleotide sequence of a cDNA for pro-ConA, and in the derived amino acid sequence the only glycosylation site is in the middle of the molecule. Furthermore, the derived amino acid sequence of the putative precursor of ConA was found not to be colinear with that of ConA. Here we show that pro-ConA is located primarily in an endoplasmic reticulum-rich organelle fraction. Pro-ConA was purified from this fraction and subjected to amino acid sequencing. The first 12 amino acids at the N-terminal end of pro-ConA correspond to amino acids 119-130 of mature ConA, and to amino acids 30-41 of the putative pre-pro-ConA, the sequence of which was derived from the nucleotide sequence of a cDNA. Amino acid sequencing of a tryptic glycopeptide with the high mannose side chain showed that the first 17 amino acids of this peptide correspond to amino acids 154-170 of pre-pro-ConA. The last six amino acids in this series correspond to the first six amino acids of mature ConA. These data fully support the hypothesis of Carrington et al. that the biosynthesis of ConA involves a post-translational peptide cleavage, transposition, and ligation within the original polypeptide. Pro-ConA from the organelle fraction does not bind to Sephadex G-50, indicating that it has no lectin activity. The processing of pro-ConA apparently imparts biological activity to this lectin.
D000595:Amino Acid Sequence; D050260:Carbohydrate Metabolism; D002846:Chromatography, Affinity; D003208:Concanavalin A / Q000032:analysis / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000032:analysis*; D007887:Fabaceae; D037121:Plant Lectins; D010944:Plants / Q000648:ultrastructure*; D010946:Plants, Medicinal; D011498:Protein Precursors / Q000032:analysis / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
D037121:Plant Lectins; D011498:Protein Precursors; D003208:Concanavalin A