scholarly journals Human Serum Albumin Binds Native Insulin and Aggregable Insulin Fragments and Inhibits Their Aggregation

Biomolecules ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1366 ◽  
Author(s):  
Joanna Wasko ◽  
Marian Wolszczak ◽  
Zbigniew J. Kaminski ◽  
Malgorzata Steblecka ◽  
Beata Kolesinska
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Butyric Acid ◽  
Hot Spots ◽  
Cd Spectra ◽  
Buffer Solution ◽  
Peptide Probe ◽  
Coupling Reagent ◽  
Native Insulin

The purpose of this study was to investigate whether Human Serum Albumin (HSA) can bind native human insulin and its A13–A19 and B12–B17 fragments, which are responsible for the aggregation of the whole hormone. To label the hormone and both hot spots, so that their binding positions within the HSA could be identified, 4-(1-pyrenyl)butyric acid was used as a fluorophore. Triazine coupling reagent was used to attach the 4-(1-pyrenyl)butyric acid to the N-terminus of the peptides. When attached to the peptides, the fluorophore showed extended fluorescence lifetimes in the excited state in the presence of HSA, compared to the samples in buffer solution. We also analyzed the interactions of unlabeled native insulin and its hot spots with HSA, using circular dichroism (CD), the microscale thermophoresis technique (MST), and three independent methods recommended for aggregating peptides. The CD spectra indicated increased amounts of the α-helical secondary structure in all analyzed samples after incubation. Moreover, for each of the two unlabeled hot spots, it was possible to determine the dissociation constant in the presence of HSA, as 14.4 µM (A13–A19) and 246 nM (B12–B17). Congo Red, Thioflavin T, and microscopy assays revealed significant differences between typical amyloids formed by the native hormone or its hot-spots and the secondary structures formed by the complexes of HSA with insulin and A13–A19 and B12–B17 fragments. All results show that the tested peptide-probe conjugates and their unlabeled analogues interact with HSA, which inhibits their aggregation.

Investigation of the Effects of Various Cyclodextrins on the Stabilisation of Human Serum Albumin by a Spectroscopic Method

2015 ◽  
Vol 68 (12) ◽  
pp. 1894 ◽  
Author(s):  
Mohsen Oftadeh ◽  
Golamreza Rezaei Behbahani ◽  
Ali Akbar Saboury ◽  
Shahnaz Rafiei
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Electrostatic Interactions ◽  
Spectroscopic Method ◽  
Phosphate Buffer Solution ◽  
Blue Shift ◽  
Titration Calorimetry ◽  
Tryptophan Residue ◽  
Buffer Solution

The binding parameters between cyclodextrins (CDs) and human serum albumin (HSA) were investigated by isothermal titration calorimetry (ITC), fluorescence quenching, and UV-vis absorption spectroscopy at 300 K in 50 mM phosphate buffer solution. Among the various CDs investigated, β-CD has the greater ability to decrease the aggregation of HSA and the results indicated that the inhibition order is γ-CD < α-CD < β-CD. The obtained heats for HSA+CDs interactions were reported and analysed in terms of the extended solvation model, which was used to reproduce the enthalpies of HSA interactions with CDs over a broad range of complex concentrations. The binding constant and thermodynamic parameters were obtained. These suggested that the binding reaction was driven by both enthalpy and entropy, and electrostatic interactions played a major role in the stabilising of HSA. The parameters and reflected the net effect of β-CD on the HSA stability at low and high cyclodextrin concentrations, respectively. The positive values for indicated that β-CD stabilises the HSA structure at low concentrations. The UV absorption intensity of theses complexes increased and a slight red shift was observed in the absorbance wavelength with increasing the CD concentration. The fluorescence intensity of HSA decreased regularly and a slight blue shift was observed for the emission wavelength with increasing CD concentration. The results indicate that the CD complex could quench the fluorescence of HSA and changes the microenvironment of the tryptophan residue.

scholarly journals Lysine residue 240 of human serum albumin is involved in high-affinity binding of bilirubin

1978 ◽  
Vol 171 (2) ◽  
pp. 453-459 ◽  
Author(s):  
C Jacobsen
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Lysine Residue ◽  
Water Soluble ◽  
High Affinity ◽  
Coupling Reagent ◽  
Peptide Fraction ◽  
Short Period ◽  
High Affinity Site

Bilirubin can be coupled covalently to albumin by using water-soluble carbodi-imide as coupling reagent. The optimal specificity in the attachment of bilirubin to the high-affinity site on the albumin molecule was obtained by treating an albumin-bilirubin complex with carbodi-imide in low concentrations and for a short period. The product was reduced, carboxymethylated and digested with trypsin. By fractionation on Sephadex G-50 (superfine grade) a peptide fraction containing most of the bilirubin label was isolated. Further purification by paper chromatography gave one peptide, consisting of residues 240-258. The peptide containined a single lysine residue, 240, and had an intact disulphide bridge. The results indicate that bilirubin is bound to lysine residue 240 at its high-affinity site on human serum albumin.

Resolution of the Absorbance and CD Spectra and Formation Constants of the Complexes between Human Serum Albumin and Methyl Orange

1996 ◽  
Vol 2 (2) ◽  
pp. 149-156 ◽  
Author(s):  
R. Ambrosetti ◽  
R. Bianchini ◽  
S. Fisichella ◽  
M. Fichera ◽  
M. Zandomeneghi
Keyword(s):  
Methyl Orange ◽  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Formation Constants ◽  
Cd Spectra

Spectroscopic Studies on the Interaction between Sulfadiazine and Human Serum Albumin

2014 ◽  
Vol 1044-1045 ◽  
pp. 181-184
Author(s):  
Fang Huang ◽  
Ying Liu
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Hydrophobic Interactions ◽  
Phosphate Buffer Solution ◽  
Inner Filter Effect ◽  
Entropy Change ◽  
Spectroscopic Studies ◽  
Fluorescence Measurement ◽  
Buffer Solution

The interaction of sulfadiazine (SDZ) and human serum albumin (HSA) in phosphate buffer solution had been investigated using multi-spectroscopic methods. The inner filter effect was corrected. The quenching mechanism was determined to be static quenching according to the fluorescence measurement. The thermodynamic parameters (enthalpy change (ΔH) and entropy change (ΔS)) were calculated to be-9.70 KJ·mol-1 and 46.07 J·mol-1·K-1, respectively, which indicated that hydrogen bonds and hydrophobic interactions play the major role on driven the interaction of SDZ with HSA. SDZ binds in the vicinity of site I in HSA, and the binding distance was 1.93 nm. In addition, the effects of HSA secondary structure were quantitatively calculated by CD spectra.

Photocyclization of photoswitches with high enantioselectivity in human serum albumin in an artificial environment

2017 ◽  
Vol 53 (22) ◽  
pp. 3181-3184 ◽  
Author(s):  
Koichi Kawamura ◽  
Ken Osawa ◽  
Yuta Watanobe ◽  
Yuri Saeki ◽  
Naoki Maruyama ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Uv Irradiation ◽  
Phosphate Buffer ◽  
Enantiomeric Excess ◽  
Phosphate Buffer Solution ◽  
Ring Closure ◽  
Buffer Solution ◽  
Artificial Environment

Three photochromic bisthienylethenes exhibited 56 to >99% enantiomeric excess in photochemical ring closure upon UV irradiation when incorporated in human serum albumin dissolved in 15% acetonitrile-phosphate buffer solution and incubated for 24 h at −4 °C.

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

1987 ◽  
Vol 65 (8) ◽  
pp. 1927-1934 ◽  
Author(s):  
M. Bouvier ◽  
G. R. Brown ◽  
L. E. St-Pierre
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Solid Phase ◽  
Solid Phase Peptide Synthesis ◽  
Phosphate Buffer Solution ◽  
Binding Constants ◽  
Buffer Solution ◽  
A Site ◽  
Scatchard Plots

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) × 104 M−1, 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.

Genetic variants showing apparent hot-spots in the human serum albumin gene

1999 ◽  
Vol 289 (1-2) ◽  
pp. 45-55 ◽  
Author(s):  
Monica Galliano ◽  
Ulrich Kragh-Hansen ◽  
András L Tárnoky ◽  
John C Chapman ◽  
Monica Campagnoli ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Genetic Variants ◽  
Hot Spots ◽  
Albumin Gene

MicroRNA 155 Downregulation by Vitamin C–Loaded Human Serum Albumin Nanoparticles During Cutaneous Wound Healing in Mice

2019 ◽  
Vol 18 (2) ◽  
pp. 143-152 ◽  
Author(s):  
Hamid Reza Shojania ◽  
Madjid Momeni-Moghaddam ◽  
Seyed-Ebrahim. Hossini ◽  
Mohammad Armin ◽  
Jalal Omrani Bidi
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Human Serum Albumin ◽  
Vitamin C ◽  
Serum Albumin ◽  
Human Serum ◽  
Control Group ◽  
Fibroblast Cells ◽  
Buffer Solution ◽  
Tgf Β1

This study focused on potential of vitamin C loaded human serum albumin (HSA) nanoparticles for treatment of wound. Nanocarrier were prepared and assessed for their effect on growth of 3T3 fibroblast cells, cell migration, wound healing rate and expression of miR-155, TGF-β1 and SMAD 1,2 genes. Wound healing assay was done and wounds were treated with vitamin C loaded HSA nanoparticles. Nanoparticles were prepared with size and zeta potential of 180±6 and -29 mV, respectively. Vitamin C loaded HSA nanoparticles showed controlled release of vitamin C into the buffer solution. Also, yield and encapsulation efficacy of loaded nanoparticles were obtained as 70.6 and 52.1 %, respectively. MTT results showed that the growth of 3T3 fibroblast cells was promoted in culture medium with 20 µg/ml of vitamin C loaded HSA nanoparticles. Cell migration assay indicated the positive effect of loaded nanoparticles on wound healing. The in-vivo results showed that the rate of wound healing was increased after treatment with 20 µg/ml of vitamin C loaded HSA nanoparticles. The wounds were healed faster when treated with vitamin C loaded HSA nanoparticles in comparison with control group. The expression of miR-155 was downregulated after treatment. Furthermore, expression of TGF-β1 and SMAD 1,2 were increased while the wounds were treated with these nanoparticles. In conclusion, these results showed for the first time that wounds were healed after treatment with albumin nanocarrier loaded with vitamin C. This nanocarrier changed expression of miR-155 and TGF-β1 towards faster healing of wounds.

A New Fluorescence Method for the Determination of Hsa Using Graphite Oxide as Nanoprobe

2013 ◽  
Vol 647 ◽  
pp. 769-773
Author(s):  
Xin Hui Zhang ◽  
Li Li Xu ◽  
Ai Hui Liang ◽  
Zhi Liang Jiang
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Ultrasonic Wave ◽  
Fluorescence Intensity ◽  
Graphite Oxide ◽  
Fluorescence Method ◽  
Buffer Solution ◽  
Fluorescence Peak

Graphite oxide (GO) was prepared by Hummer procedure, and can be dispersed to obtain stable GO nanocolloid solution by ultrasonic wave. The GO exhibited a weak fluorescence peak at 425 nm in pH 4.6 HAc-NaAc buffer solution. Upon addition of human serum albumin (HSA), it combined with GO nanoprobe to form big HSA-GO particles that caused the fluorescence peak increasing. The increased fluorescence intensity was linear to HSA concentration in the range of 2-200 μg/mL. Thus, a new and simple fluorescence method was proposed for the determination of HSA in real sample.

Multivariate Determination of Human Serum Albumin and γ-Globulin in a Phosphate Buffer Solution by near Infrared Spectroscopy

1998 ◽  
Vol 6 (1) ◽  
pp. 375-381 ◽  
Author(s):  
Koichi Murayama ◽  
Keiichi Yamada ◽  
Roumiana Tsenkova ◽  
Yan Wang ◽  
Yukihiro Ozaki
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Standard Error ◽  
Phosphate Buffer ◽  
Near Infrared ◽  
Phosphate Buffer Solution ◽  
Clinical Analysis ◽  
Pls Regression ◽  
Buffer Solution

Near infrared (NIR) spectra in the 1300–1850 nm region were measured for phosphate buffer solutions containing both human serum albumin and γ-globulin with various concentrations. The concentrations of albumin and γ-globulin were determined by partial least squares (PLS) regression analysis. The calibration for albumin in the concentration ranges of 0.100–8.000 g dL−1 yielded the correlation coefficient ( R) of 0.998, the standard error of calibration ( SEC) of 0.124 g dL−1, and the standard error of prediction ( SEP) of 0.152 g dL−1, respectively. Those for γ-globulin in the concentration ranges of 0.100–6.000 g dL−1 yielded were 0.998, 0.110 g dL−1 and 0.118 g dL−1, respectively. The regression coefficients ( RCs) of PLS factors for albumin were compared with those for γ-globulin. The differences were discussed for each RC between albumin and γ-globulin. The coefficient of variation ( CV) was calculated to be 0.0388 and 0.0374 for albumin and γ-globulin, respectively. The ratio of standard derivation of reference data in prediction set to SEP ( RPD) are 13.4 and 16.4 for them. These values obtained in the present study satisfy the demands of clinical analysis of blood. The present results demonstrate that it is possible to determine the concentrations of the two kinds of proteins in the solution simultaneously by use of NIR and PLS regression.

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