Evidence for a single naphthylphthalamic acid binding site on the zucchini plasma membrane

The binding of [2,3,4,5,(n)-3H]N-1-napthylphthalamic acid ([3H]-NPA) to zucchini (Cucurbita pepo L.) plasma membranes was examined in detail using two different filtration assays and the results were rigorously analyzed by saturation curves, double-reciprocal plots, Scatchard plots, Hill plots, and...

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Bibliographic Details
Published in:Plant physiology (Bethesda) 1993-10, Vol.103 (2), p.449-456
Main Authors: Muday, G.K, Brunn, S.A, Haworth, P, Subramanian, M
Format: Article
Language:English
Subjects:
ACIDE ORGANIQUE
ACIDOS ORGANICOS
ANALISIS CUANTITATIVO
ANALYSE QUANTITATIVE
Auxins
Binding sites
Biological and medical sciences
Cell membranes
Cell receptors
Cell structures and functions
Centrifugation
Coleoptiles
Corn
CUCURBITA PEPO
DESINFECTANT
DESINFECTANTES
ESTRUCTURA CELULAR
FILTRE
FILTROS
Fundamental and applied biological sciences. Psychology
Growth regulators
Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors
Hypocotyls
Kinetics
Ligands
Metabolism
Metabolism and Enzymology
Molecular and cellular biology
ORGANITE CELLULAIRE
ORGANULOS CITOPLASMICOS
Plant physiology and development
Receptors
STRUCTURE CELLULAIRE
ULTRAFILTRACION
ULTRAFILTRATION
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Summary:The binding of [2,3,4,5,(n)-3H]N-1-napthylphthalamic acid ([3H]-NPA) to zucchini (Cucurbita pepo L.) plasma membranes was examined in detail using two different filtration assays and the results were rigorously analyzed by saturation curves, double-reciprocal plots, Scatchard plots, Hill plots, and the computer program Ligand (P.J. Munson, D. Rodbard [1980] Anal Biochem 107:220-239). To facilitate these analyses, a new assay that allows rapid and quantitative analysis of [3H]NPA binding with high reproducibility and ease of manipulation has been developed. These detailed kinetic analyses indicate that only one binding site for [3H]NPA (Kd = 16 nM) was associated with the zucchini plasma membrane. Analysis of [3H]NPA dissociation by several auxin transport inhibitors revealed similar dissociation constants with both plasma and microsomal membrane. Collectively, these data indicate the presence of only one binding site for NPA associated with the zucchini plasma membrane
ISSN:0032-0889
1532-2548
DOI:10.1104/pp.103.2.449