Solubilisation and partial purification of auxin-binding sites of corn membranes

Mathematics – Logic

Scientific paper

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Scientific paper

THE demonstration by Hertel et al.1 of reversible, high-affinity binding of auxins to membranous preparations from corn coleoptiles provided for the first time a reliable and readily accessible system for the study of potential receptor-phytohormone interactions. In a detailed investigation of the binding kinetics and specificity of this system, we presented evidence for the presence of two classes of auxin-binding sites with differing affinities and specificities2,3. Site 1, with a KD for the synthetic auxin 1-naphthalene-acetic acid (NAA) of 0.14 µM, was able to bind both auxins and physiologically inactive analogues, whereas site (KD for NAA = 1.7 µM) displayed binding specificity compatible with that expected of an auxin receptor2 and was located in a heavier membrane population, enriched in plasma membrane3. Auxin-binding activity can be readily solubilised from the membranes with Triton X-100 (ref. 2), but our attempts to purify such extracts further have met with only limited success. I report here that both binding sites can be conveniently removed from the membranes without detergent and purified approximately 100-fold in two steps.

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