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Related Experiment Videos

Weak base binding and transport in pig brush border membrane vesicles.

A J Moe1, J A Hollywood, M J Jackson

  • 1George Washington University Medical Center, Washington, District of Columbia.

The Journal of Pharmacology and Experimental Therapeutics
|September 1, 1988
PubMed
Summary

Weak bases like benzylamine bind to pig intestinal membranes, suggesting specific binding sites for non-ionized amines. This binding may influence how these compounds are transported across the intestinal barrier.

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Area of Science:

  • Biochemistry
  • Physiology
  • Pharmacology

Background:

  • Weak bases are absorbed in the intestine, but the mechanisms are not fully understood.
  • Intestinal brush border membranes play a crucial role in nutrient and drug absorption.
  • Understanding the interaction of weak bases with these membranes is key to predicting their bioavailability.

Purpose of the Study:

  • To characterize the uptake and binding of weak bases, specifically benzylamine and amphetamine, in pig intestinal brush border membrane vesicles.
  • To investigate the role of pH and ionic strength on weak base binding.
  • To determine the kinetic and structural properties of these binding interactions.

Main Methods:

  • Utilized pig intestinal brush border membrane vesicles to study weak base uptake.

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  • Assessed osmotic sensitivity to differentiate binding from transport.
  • Performed pH-dependent binding studies and kinetic analyses (activation energy, saturation kinetics).
  • Conducted inhibition experiments with structurally related compounds.

    • Main Results:

    • Weak bases showed minimal transport but significant binding at physiological pH (7.4).
    • Benzylamine binding increased with pH, indicating preference for the non-ionized form.
    • Binding was saturable, with two distinct binding sites identified (KD values of 25 µM and 2.5 mM).
    • High structural selectivity and evidence of hydrogen bond acceptor interactions were observed.

    Conclusions:

    • Non-ionized primary amines bind to specific sites on intestinal brush border membranes.
    • This specific binding suggests a role in the translocation or membrane discrimination of weak bases.
    • The findings provide insights into the molecular interactions governing weak base absorption in the intestine.