Location and characterization of the warfarin binding site of human serum albumin

Bos, Octaaf J.M.; Remijn, Jobina P.M.; Fischer, Marcel J.E.; Wilting, Jaap; Janssen, Lambert H.M.

doi:10.1016/0006-2952(88)90072-x

Cited by 83 publications

(38 citation statements)

References 18 publications

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“…Fragments of HSA produced by chemical or enzymatic cleavage have been used to define the exact high-affinity binding sites for several ligands [7][8][9][10] . However, this method is limited by the finite number of cleavage sites in HSA, and chemical cleavage may destroy the structure of the binding site.…”

Section: Introductionmentioning

confidence: 99%

Stereoselective binding of mexiletine and ketoprofen enantiomers with human serum albumin domains

et al. 2012

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Aim: To investigate the stereoselective binding of mexiletine or ketoprofen enantiomers with different recombinant domains of human serum albumin (HSA). Methods: Three domains (HSA DOM I, II and III) were expressed in Pichia pastoris GS115 cells. Blue Sepharose 6 Fast Flow was employed to purify the recombinant HSA domains. The binding properties of the standard ligands, digitoxin, phenylbutazone and diazepam, and the chiral drugs to HSA domains were investigated using ultrafiltration. The concentrations of the standard ligands, ketoprofen and mexiletine were analyzed with HPLC. Results: The recombinant HSA domains were highly purified as shown by SDS-PAGE and Western blotting analyses. The standard HSA ligands digitoxin, phenylbutazone and diazepam selectively binds to DOM I, DOM II and DOM III, respectively. For the chiral drugs, R-ketoprofen showed a higher binding affinity toward DOM III than S-ketoprofen, whereas S-mexiletine bound to DOM II with a greater affinity than R-mexiletine. Conclusion:The results demonstrate that HSA DOM III possesses the chiral recognition ability for the ketoprofen enantiomers, whereas HSA DOM II possesses that for the mexiletine enantiomers.

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Section: Introductionmentioning

confidence: 99%

Stereoselective binding of mexiletine and ketoprofen enantiomers with human serum albumin domains

et al. 2012

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“…The precise nature of the structural changes associated with the neutral-basic transition is incompletely characterized. The transition appears largely to involve changes in domains I and II, although there is some involvement of domain III (32). It remains unclear whether there is any link between the structural changes induced by shifts in pH and those induced by fatty acid binding, although the observation that fatty acid binding lowers the pH of the midpoint of the neutral-basic transition suggests a possible parallel between the two effects (25).…”

Section: Discussionmentioning

confidence: 99%

Crystal Structure Analysis of Warfarin Binding to Human Serum Albumin

Petitpas¹,

Bhattacharya²,

Twine³

et al. 2001

Journal of Biological Chemistry

777

605

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Human serum albumin (HSA) is an abundant transport protein found in plasma that binds a wide variety of drugs in two primary binding sites (I and II) and can have a significant impact on their pharmacokinetics. We have determined the crystal structures at 2.5 Å-resolution of HSA-myristate complexed with the R-(؉) and S-(؊) enantiomers of warfarin, a widely used anticoagulant that binds to the protein with high affinity. The structures confirm that warfarin binds to drug site I (in subdomain IIA) in the presence of fatty acids and reveal the molecular details of the protein-drug interaction. The two enantiomers of warfarin adopt very similar conformations when bound to the protein and make many of the same specific contacts with amino acid side chains at the binding site, thus accounting for the relative lack of stereospecificity of the HSA-warfarin interaction. The conformation of the warfarin binding pocket is significantly altered upon binding of fatty acids, and this can explain the observed enhancement of warfarin binding to HSA at low levels of fatty acid.

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“…Protein concentrations were measured by their absorbance at 278 nm on a Hewlett-Packard HP8453 spectrophotometer. For HSA, an absorption coefficient (A 278 nm,1 cm 0.1% ) of 0.58 was used (8). For HSA-DOM I, HSA-DOM II, and HSA-DOM III, the absorption coefficients were found to be 0.50, 0.79, and 0.30, respectively (3).…”

Section: Methodsmentioning

confidence: 99%

Conformational Transitions of the Three Recombinant Domains of Human Serum Albumin Depending on pH

Dockal¹,

Carter²,

Rüker³

2000

Journal of Biological Chemistry

431

323

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Human serum albumin (HSA) is a protein of 66.5 kDa that is composed of three homologous domains, each of which displays specific structural and functional characteristics. HSA is known to undergo different pH-dependent structural transitions, the N-F and F-E transitions in the acid pH region and the N-B transition at slightly alkaline pH. In order to elucidate the structural behavior of the recombinant HSA domains as standalone proteins and to investigate the molecular and structural origins of the pH-induced conformational changes of the intact molecule, we have employed fluorescence and circular dichroic methods. Here we provide evidence that the loosening of the HSA structure in the N-F transition takes place primarily in HSA-DOM III and that HSA-DOM I undergoes a structural rearrangement with only minor changes in secondary structure, whereas HSA-DOM II transforms to a molten globulelike state as the pH is reduced. In the pH region of the N-B transition of HSA, HSA-DOM I and HSA-DOM II experience a tertiary structural isomerization, whereas with HSA-DOM III no alterations in tertiary structure are observed, as judged from near-UV CD and fluorescence measurements.

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Location and characterization of the warfarin binding site of human serum albumin

Cited by 83 publications

References 18 publications

Stereoselective binding of mexiletine and ketoprofen enantiomers with human serum albumin domains

Stereoselective binding of mexiletine and ketoprofen enantiomers with human serum albumin domains

Crystal Structure Analysis of Warfarin Binding to Human Serum Albumin

Conformational Transitions of the Three Recombinant Domains of Human Serum Albumin Depending on pH

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