N-1-napthylphthalamic-acid-binding activity of a plasma membrane-rich fraction from maize coleoptiles

Abstract

Plasma membrane-rich fractions were prepared from maize coleoptiles by low-shear homogenization and differential and sucrose-gradient centrifugation. Plasma membrane fragments were identified using a specific cytochemical stain based on phosphotungstic acid prepared in chromic acid. In a comparison of 10 different cell fractions of varying plasma membrane content, the N-1-napthylphthalamic-acid (NPA)-binding activity of the fractions was directly proportional to the content of plasma membrane. The NPA binding appears to be strong K_M between 10^-8 and 10^-7 M) but non-covalent. NPA is known to inhibit auxin transport efficiently and quickly. Thus, the results are consistent with the localization of auxin transport sites at the plasma membrane of plant cells.