Binding of [3H]Imipramine to Human Platelet Membranes with Compensation for Saturable Binding to Filters and Its Implication for Binding Studies with Brain Membranes

doi:10.1111/j.1471-4159.1984.tb00924.x

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Wiley InterScience

Journal of Neurochemistry

Volume 43 Issue 2, Pages 479 - 486

Published Online: 5 Oct 2006

Published for the International Society for Neurochemistry

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Binding of [³H]Imipramine to Human Platelet Membranes with Compensation for Saturable Binding to Filters and Its Implication for Binding Studies with Brain Membranes

Orla M. Phillips,* Keith M. Wood,† D. Clive Williams*

*Department of Biochemistry, Trinity College, Dublin, Ireland †M.R.C. Neuropsychiatry Laboratory, West Park Hospital, Epsom, Surrey, U.K.

Address correspondence and reprint requests to D. Clive Williams, Department of Biochemistry, Trinity College, Dublin 2, Ireland.

KEYWORDS

Imipramine • Human platelets • Saturable filter binding

ABSTRACT

Abstract: Apparent specific binding of [³H]imipramine to human platelet membranes at high concentrations of imipramine showed deviation from that expected of a single binding site, a result consistent with a low-affinity binding site. The deviation was due to displaceable, saturable binding to the glass fibre filters used in the assays. Imipramine, chloripramine, desipramine, and fluoxetine inhibited binding to filters whereas 5-hydroxytryptamine and ethanol were ineffective. Experimental conditions were developed that eliminated filter binding, allowing assay of high and low-affinity binding to membranes. Failure to correct for filter binding may lead to overestimation of binding parameters, B_max and K_D for high-affinity binding to membranes, and may also be misinterpreted as indicating a low-affinity binding component in both platelet and brain membranes. Low-affinity binding (K_D < 2 μM) of imipramine to human platelet membranes was demonstrated and its significance discussed.

Received December 9, 1983; accepted January 25, 1984.

DIGITAL OBJECT IDENTIFIER (DOI)

10.1111/j.1471-4159.1984.tb00924.x About DOI