Studies on the biotin-binding site of avidin. Tryptophan residues involved in the active site.
Egg-white avidin was modified with the tryptophan-specific reagent 2-hydroxy-5-nitrobenzyl bromide. The complete loss of biotin-binding activity was achieved upon modification of an average of one tryptophan residue per avidin subunit. The identity of the modified residues was determined by isolating the relevant tryptic and chymotryptic peptides from CNBr-cleaved avidin fragments. The results demonstrate that Trp-70 and Trp-110 are modified in approximately equivalent proportions. It is believed that these residues are located in the active site of avidin and take part in the binding of biotin.
Gitlin
G
G
Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Bayer
E A
EA
Wilchek
M
M
D000596:Amino Acids / Q000032:analysis; D001360:Avidin / Q000378:metabolism*; D001665:Binding Sites; D001710:Biotin / Q000378:metabolism*; D002850:Chromatography, Gel; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D014364:Tryptophan / Q000032:analysis*
D000596:Amino Acids; D010446:Peptide Fragments; D001360:Avidin; D001710:Biotin; D014364:Tryptophan