Determining molecular binding sites on human serum albumin by displacement of oleic acid

Ronald W. Sarver, Hua Gao, Fang Tian

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

An NMR method was developed for determining binding sites of small molecules on human serum albumin (HSA) by competitive displacement of 13C-labeled oleic acid. This method is based on the observation that in the crystal structure of HSA complexed with oleic acid, two principal drug-binding sites, Sudlow's sites I (warfarin) and II (ibuprofen), are also occupied by fatty acids. In two-dimensional [1H,13C] heteronuclear single quantum coherence NMR spectra, seven distinct resonances were observed for the 13C-methyl-labeled oleic acid as a result of its binding to HSA. Resonances corresponding to the major drug-binding sites were identified through competitive displacement of molecules that bind specifically to each site. Thus, binding of molecules to these sites can be followed by their displacement of oleic acids. Furthermore, the amount of bound ligand at each site can be determined from changes in resonance intensities. For molecules containing fluorine, binding results were further validated by direct observations of the bound ligands using 19F NMR. Identifying the binding sites for drug molecules on HSA can aid in determining the structure-activity relationship of albumin binding and assist in the design of molecules with altered albumin binding.

Original languageEnglish (US)
Pages (from-to)297-302
Number of pages6
JournalAnalytical Biochemistry
Volume347
Issue number2
DOIs
StatePublished - Dec 15 2005

Fingerprint

Oleic Acid
Serum Albumin
Binding Sites
Molecules
Albumins
Nuclear magnetic resonance
Pharmaceutical Preparations
Oleic Acids
Ligands
Fluorine
Ibuprofen
Warfarin
Structure-Activity Relationship
Fatty Acids
Crystal structure

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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abstract = "An NMR method was developed for determining binding sites of small molecules on human serum albumin (HSA) by competitive displacement of 13C-labeled oleic acid. This method is based on the observation that in the crystal structure of HSA complexed with oleic acid, two principal drug-binding sites, Sudlow's sites I (warfarin) and II (ibuprofen), are also occupied by fatty acids. In two-dimensional [1H,13C] heteronuclear single quantum coherence NMR spectra, seven distinct resonances were observed for the 13C-methyl-labeled oleic acid as a result of its binding to HSA. Resonances corresponding to the major drug-binding sites were identified through competitive displacement of molecules that bind specifically to each site. Thus, binding of molecules to these sites can be followed by their displacement of oleic acids. Furthermore, the amount of bound ligand at each site can be determined from changes in resonance intensities. For molecules containing fluorine, binding results were further validated by direct observations of the bound ligands using 19F NMR. Identifying the binding sites for drug molecules on HSA can aid in determining the structure-activity relationship of albumin binding and assist in the design of molecules with altered albumin binding.",
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Determining molecular binding sites on human serum albumin by displacement of oleic acid. / Sarver, Ronald W.; Gao, Hua; Tian, Fang.

In: Analytical Biochemistry, Vol. 347, No. 2, 15.12.2005, p. 297-302.

Research output: Contribution to journalArticle

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