Affinity labels for auxin binding sites in corn coleoptile membranes

Two auxin analogues have been tested as affinity labels for auxin binding sites in coleoptile membranes of Zea mays L. Reacting the membranes at pH 8—9 with the diazonium salt of CAPA (2-chloro-4-aminophenoxyacetic acid) reduces their subsequent ability to bind NAA(1-naphthylacetic acid). Diazo-Chloramben (2,5-dichloro-3-aminobenzoic acid) is also effective in inhibiting NAA binding capacity and this inhibition is largely independent of reaction pH over the range pH 6—9. Similar experiments with sulphydryl reagents have shown that reaction of the membranes with p-mercuribenzoate (PMB) strongly inhibits subsequent auxin binding activity. Prior addition of NAA protects the binding sites against the action of diazo-Chloramben or PMB when the reactions are carried out at pH 6. From these results and from other considerations, several of the amino acid residues in the binding site environment have been tentatively assigned.