Characterization of thyroxine-albumin binding using high-performance affinity chromatography. I. Interactions at the warfarin and indole sites of albumin

Bounthon Loun, David S. Hage

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110 Citations (Scopus)

Abstract

A high-performance affinity column containing immobilized human serum albumin (HSA) was used to study the binding of thyroxine at the warfarin and indole sites of HSA. Frontal analysis, using R-warfarin and l-tryptophan as probes for these sites, demonstrated that the immobilized HSA had binding behavior equivalent to that observed for HSA in solution. By injecting R-warfarin or l-tryptophan in the presence of excess thyroxine, it was found that thyroxine was binding directly to both types of site. The warfarin and indole sites had relatively strong binding for thyroxine, with association constants at 37°C of 1.4 · 105 and 5.7 · 105 M-1, respectively. The value of δG for these sites ranged from -7 to -8 kcal/mol and had a significant entropy component. The techniques used in this study are not limited to thyroxine-HSA interactions, but should also be valuable in examining the site-specific binding of other drugs and hormones to HSA.

Original languageEnglish (US)
Pages (from-to)225-235
Number of pages11
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume579
Issue number2
DOIs
StatePublished - Sep 4 1992

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Affinity chromatography
Warfarin
Thyroxine
Affinity Chromatography
Serum Albumin
Albumins
Tryptophan
Entropy
indole
Binding Sites
Association reactions
Hormones
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

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abstract = "A high-performance affinity column containing immobilized human serum albumin (HSA) was used to study the binding of thyroxine at the warfarin and indole sites of HSA. Frontal analysis, using R-warfarin and l-tryptophan as probes for these sites, demonstrated that the immobilized HSA had binding behavior equivalent to that observed for HSA in solution. By injecting R-warfarin or l-tryptophan in the presence of excess thyroxine, it was found that thyroxine was binding directly to both types of site. The warfarin and indole sites had relatively strong binding for thyroxine, with association constants at 37°C of 1.4 · 105 and 5.7 · 105 M-1, respectively. The value of δG for these sites ranged from -7 to -8 kcal/mol and had a significant entropy component. The techniques used in this study are not limited to thyroxine-HSA interactions, but should also be valuable in examining the site-specific binding of other drugs and hormones to HSA.",
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