Isolation and identification of a trypsin-resistant fragment of human serum albumin with bilirubin- and drug-binding properties

Sjödin, Torgny; Hansson, Ronnie; Sjöholm, Ingvar

doi:10.1016/0005-2795(77)90135-0

Cited by 30 publications

(14 citation statements)

References 45 publications

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“…This is in accordance with previous findings that the binding of some ligands to HSA, such as anionic azo-dyes [38] and bilirubin [39], a structural analog of PCB, decreases the susceptibility of HSA to hydrolytic attack by trypsin. Increased compactness and reduced flexibility of HSA induced by PCB binding protected the protein from proteolytic degradation, as susceptibility to proteolysis is determined by exposure of peptide bonds, as well as by mobility of protein segments containing these bonds.…”

Section: Discussionsupporting

confidence: 93%

Stabilization of Human Serum Albumin by the Binding of Phycocyanobilin, a Bioactive Chromophore of Blue-Green Alga Spirulina: Molecular Dynamics and Experimental Study

et al. 2016

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Phycocyanobilin (PCB) binds with high affinity (2.2 x 106 M-1 at 25°C) to human serum albumin (HSA) at sites located in IB and IIA subdomains. The aim of this study was to examine effects of PCB binding on protein conformation and stability. Using 300 ns molecular dynamics (MD) simulations, UV-VIS spectrophotometry, CD, FT-IR, spectrofluorimetry, thermal denaturation and susceptibility to trypsin digestion, we studied the effects of PCB binding on the stability and rigidity of HSA, as well as the conformational changes in PCB itself upon binding to the protein. MD simulation results demonstrated that HSA with PCB bound at any of the two sites showed greater rigidity and lower overall and individual domain flexibility compared to free HSA. Experimental data demonstrated an increase in the α-helical content of the protein and thermal and proteolytic stability upon ligand binding. PCB bound to HSA undergoes a conformational change to a more elongated conformation in the binding pockets of HSA. PCB binding to HSA stabilizes the structure of this flexible transport protein, making it more thermostable and resistant to proteolysis. The results from this work explain at molecular level, conformational changes and stabilization of HSA structure upon ligand binding. The resultant increased thermal and proteolytic stability of HSA may provide greater longevity to HSA in plasma.

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Section: Discussionsupporting

confidence: 93%

Stabilization of Human Serum Albumin by the Binding of Phycocyanobilin, a Bioactive Chromophore of Blue-Green Alga Spirulina: Molecular Dynamics and Experimental Study

et al. 2016

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“…Drug binding studies have for a long time relied on the use of albumin fragments, which proved to be valuable research tools, allowing the dissection and separation of the complex array of different binding sites present on HSA from each other, thereby facilitating the study of the binding of ligands that have more than one binding site on albumin (10,12,13,25,33,34).…”

Section: Discussionmentioning

confidence: 99%

The Three Recombinant Domains of Human Serum Albumin

Dockal¹,

Carter

Rüker³

1999

Journal of Biological Chemistry

379

184

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In an attempt to systematically dissect the ligand binding properties of human serum albumin (HSA), the gene segments encoding each of its three domains were defined based on their conserved homologous structural motifs and separately cloned into a secretion vector for Pichia pastoris. We were able to establish a generally applicable purification protocol based on Cibacron Blue affinity chromatography, suggesting that each of the three domains carries a binding site specific for this ligand. Proteins were characterized by SDS-polyacrylamide gel electrophoresis, isoelectric focusing, gel filtration, N-terminal sequencing, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, as well as near-and far-UV CD. In addition to the affinity chromatography ligand Cibacron Blue, binding properties toward hemin, warfarin, and diazepam, each of which represents a standard ligand for HSA, respectively, were investigated by the measurement of induced circular dichroism. Clear experimental evidence is provided here for the location of the primary hemin binding site to be on domain I of HSA, and for the primary diazepam binding site to be on domain III. Further, secondary binding sites were found for hemin to be located on domains II and III, and for diazepam on domain I. The warfarin binding site was located primarily on domain II, while on domain I, a secondary binding site and/or parts of the primary binding site were found.

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“…With knowledge of the primary structure of HSA (7,8) and information available concerning the possible location of the binding sites for bilirubin on that molecule (9)(10)(11)(12)(13), it is possible to synthesize onto a suitable polymer support peptide sequences that are replicates of corresponding sections of the primary structure of H S A and to test these materials as adsorbents for bilirubin.…”

Section: Introductionmentioning

confidence: 99%

Immobilized replicates of sequence 136–148 of human serum albumin as adsorbents for bilirubin

1987

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. Can. J. Chem. 65, 1927Chem. 65, (1987. Strategic peptide sequences, patterned on the sequence 136-148 of the primary structure of human serum albumin, have been immobilized on a cross-linked polyacrylamide support using the solid phase peptide synthesis technique. Certain of the resulting materials proved to be efficient adsorbents for bilirubin from aqueous phosphate buffer solution. Amino acids such as lysine and arginine favour the binding of the ligand, whereas glutamic acid reduces it markedly. From Scatchard plots, first and second equilibrium binding constants, in the range of (0.3-9.6) x lo4 M-I, were obtained using a site treatment. These binding constants are comparable to that for the binding of bilirubin by a larger fragment of bovine serum albumin that contains the synthesized sequence. Chem. 65, 1927Chem. 65, (1987.Des chaines peptidiques identiques a la sequence 136-148 de la structure primaire de l'albumine humaine ont Ct C immobilisCes sur un support de polyacrylamide rCticulC en utilisant la synthkse en phase solide des peptides. Certains des polymkres ii contenu peptidique ainsi obtenus se sont avert% Ctre d'excellents adsorbants pour la bilirubine lorsque testes en milieu tampon. La presence d'acides aminks tels que I'arginine et la lysine dans la sequence peptidique favorisent l'adsorption du ligand tandis que I'acide glutamique la rCduit apprkciablement. Des constantes d'associations de l'ordre de (0,3-9,6) X lo4 M-' ont Ct C obtenues pour ces adsorbants directement des courbes de Scatchard ii I'aide d'un traitement mathkmatique bask sur les sites d'adsorption. Ces constantes d'associations sont comparables a celle pour l'association de la bilirubine sur un fragment de l'albumine bovine qui contient la sequence synthetisee.

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Isolation and identification of a trypsin-resistant fragment of human serum albumin with bilirubin- and drug-binding properties

Cited by 30 publications

References 45 publications

Stabilization of Human Serum Albumin by the Binding of Phycocyanobilin, a Bioactive Chromophore of Blue-Green Alga Spirulina: Molecular Dynamics and Experimental Study

Stabilization of Human Serum Albumin by the Binding of Phycocyanobilin, a Bioactive Chromophore of Blue-Green Alga Spirulina: Molecular Dynamics and Experimental Study

The Three Recombinant Domains of Human Serum Albumin

Immobilized replicates of sequence 136–148 of human serum albumin as adsorbents for bilirubin

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