Photoaffinity labeling of the adenine binding site of the lectins from lima bean, Phaseolus lunatus, and the kidney bean, Phaseolus vulgaris.
8-Azidoadenine was employed as a photoaffinity probe of the adenine binding site of the seed lectin from lima beans and from Phaseolus vulgaris erythroagglutinin. This compound was shown to (a) bind competitively to the adenine binding site of these lectins and (b) exhibit enhanced binding in the presence of 1,8-anilinonaphthalenesulfonic acid in the same manner as adenine. The presence or absence of 1,8-anilinonaphthalenesulfonic acid during labeling caused no change in the peptide maps of either lectin when digested with trypsin. The peptide maps of each lectin showed one major peak of radioactivity. Sequencing of the corresponding tryptic peptide from lima bean lectin indicated the primary structure to be Val-Leu-Ile-Thr-Tyr-Asp-Ser-Ser-Thr-Lys. The sequence of the labeled peptide isolated from P. vulgaris erythroagglutinin was Thr-Thr-Thr-Trp-Asp-Phe-Val-Gly-Glu-Asn-Glu-Val-Leu-Ile-Thr-Tyr, which corresponded to residues 173-190 of the cDNA-derived sequence (Hoffman, L. M., and Donaldson, D. D. (1985) EMBO J. 4, 883-889). Residues 186-190 (italicized) are identical to the first five amino acids in the lima bean lectin peptide. The peptides are located at the COOH-terminal half of the lectin and show extensive homology with other legume lectins.
Maliarik
M J
MJ
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109.
Goldstein
I J
IJ
D000225:Adenine / Q000031:analogs & derivatives* / Q000378:metabolism*; D000345:Affinity Labels / Q000378:metabolism*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001386:Azides / Q000378:metabolism*; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D037102:Lectins / Q000378:metabolism*; D010777:Photochemistry; D010835:Phytohemagglutinins / Q000378:metabolism*; D037121:Plant Lectins
D000345:Affinity Labels; D000596:Amino Acids; D001386:Azides; D037102:Lectins; D010835:Phytohemagglutinins; D037121:Plant Lectins; C031887:lima bean lectin; C032235:8-azidoadenine; D000225:Adenine