Interaction of human serum albumin with oxovanadium ions studied by FT-IR spectroscopy and gel and capillary electrophoresis

2001 ◽  
Vol 79 (10) ◽  
pp. 1415-1421 ◽  
Author(s):  
M Purcell ◽  
J F Neault ◽  
H Malonga ◽  
H Arakawa ◽  
H A Tajmir-Riahi
Keyword(s):  
Capillary Electrophoresis ◽  
Human Serum Albumin ◽  
Ir Spectroscopy ◽  
Metal Ion ◽  
Binding Mode ◽  
Ion Binding ◽  
Metal Ion Binding ◽  
Ft Ir ◽  
Ft Ir Spectroscopy ◽  
Oxovanadium Compounds

Some oxovanadium compounds have shown potential to inhibit RNase activity, while at the same time not inhibiting DNase activity. Some vanadyl complexes also inhibit protein synthesis in rabbit reticulocytes, but induce activation of protein–tyrosine kinase. To gain an insight into the interaction of oxovanadium ions with proteins, the present study was designed to examine the bindings of VOSO4 and NaVO3 salts with human serum albumin (HSA) in aqueous solution at physiological pH with metal ion concentrations of 0.0001 to 1 mM and HSA (fatty acid free) concentration of 2% w/v. Gel and capillary electrophoresis (CE) and Fourier transform infrared (FT-IR) spectroscopic methods were used to determine the metal ion binding mode, association constant, and the secondary structure of the protein in the presence of the oxovanadium compounds. Gel electrophoresis results showed that a maximum of 20 vanadyl cations (VO2+) are bound per HSA molecule with strong (K1 = 7.0 × 107 M–1) and weak (K2 = 6.5 × 105 M–1) bindings. Similarly, capillary electrophoresis showed two major bindings for vanadyl cation with K1 = 1.2 × 108 M–1 and K2 = 8.5 × 105 M–1, whereas vanadate (VO–3) has only a weak binding affinity (K = 6.0 × 103 M–1) with HSA molecule. The VO–3 binds mainly to the lysine ε-amino NH+3 groups, while VO2+ binds possibly to the histidine nitrogen atom and the N-terminal of the α-amine residue. Infrared spectroscopic analysis showed metal ion binding results in major protein secondary structural changes from that of the α-helix (55.0 to 43–44%) to the β-sheet (22.0 to 23–26%), β-antiparallel (12.0 to 13–16%), and turn (11.0 to 17–18%), at high metal ion concentration. The observed spectral changes indicate a partial unfolding of the protein structure, in the presence of oxovanadium ions.Key words: oxovanadium, protein, binding mode, binding constant, secondary structure, electrophoresis, FT-IR spectroscopy.

Interaction of RNase A with VO3-and VO2+Ions. Metal Ion Binding Mode and Protein Secondary Structure

1999 ◽  
Vol 17 (3) ◽  
pp. 473-480 ◽  
Author(s):  
M. Purcell ◽  
A. Novetta-Delen ◽  
H. Arakawa ◽  
H. Malonga ◽  
H. A. Tajmir-Riahi
Keyword(s):  
Secondary Structure ◽  
Metal Ion ◽  
Protein Secondary Structure ◽  
Binding Mode ◽  
Rnase A ◽  
Ion Binding ◽  
Metal Ion Binding

SPECTROSCOPIC STUDY OF PROPOFOL BINDING TO HUMAN SERUM ALBUMIN

2010 ◽  
Vol 05 (04) ◽  
pp. 209-226 ◽  
Author(s):  
SAQER M. DARWISH
Keyword(s):  
Human Serum Albumin ◽  
Ir Spectroscopy ◽  
Serum Albumin ◽  
Human Serum ◽  
Protein Secondary Structure ◽  
Complex Molecules ◽  
Ft Ir ◽  
Β Sheets ◽  
Α Helix ◽  
Ft Ir Spectroscopy

The interaction of propofol and human serum albumin (HSA) has been investigated by UV-absorption, fluorescence spectroscopy and Fourier transform infrared (FT-IR) spectroscopy. Propofol has shown a strong ability to quench the intrinsic fluorescence of HSA through a static quenching procedure. The binding constant (k) is estimated at a low value of 2.55 × 103M-1at 293 K. FT-IR spectroscopy with Fourier self-deconvolution technique was used to determine the protein secondary structure in the amide regions I, II and III. The observed spectral changes of HSA-propofol complex indicate a larger intensity decrease in the absorption band of α-helix relative to that of β-sheets. This variation in intensity is related indirectly to the formation of H-bonding in the complex molecules, which accounts for the different intrinsic propensities of α-helix and β-sheets.

Chromium (III) thiamin diphosphate: a new probe of enzyme coenzyme–metal ion binding. Interaction with wheat germ pyruvate decarboxylase

1978 ◽  
Vol 56 (10) ◽  
pp. 999-1001 ◽  
Author(s):  
Ronald Kluger ◽  
Timothy Smyth
Keyword(s):  
Metal Ion ◽  
Wheat Germ ◽  
Pyruvate Decarboxylase ◽  
Binding Mode ◽  
Exchange Chromatography ◽  
Ion Binding ◽  
Specific Information ◽  
Metal Ion Binding ◽  
Thiamin Diphosphate ◽  
Binding Interaction

Chromium (III) thiamin diphosphate (CrTDP) is a substitutionally inert complex which is a physical and kinetic probe of the binding mode of metal ion and coenzyme in thiamin diphosphate (TDP) dependent enzymes. CrTDP is prepared by reaction of aquated Cr (III) and TDP and purified by ion-exchange chromatography. CrTDP binds to the apoenzyme of wheat germ pyruvate decarboxylase, giving an inactive holoenzyme. Chromium (III) ion binds to the apoenzyme in a manner that suggests, in light of the results with CrTDP, that there are two metal ion binding sites. Extension of the use of CrTDP to other enzymes can give specific information about binding and can introduce an active site reporter group.

Monitoring Metal Ion Binding in Single-LayerPseudomonas aeruginosaBiofilms Using ATR−IR Spectroscopy

Langmuir ◽  
2006 ◽  
Vol 22 (1) ◽  
pp. 286-291 ◽  
Author(s):  
So-Young Kang ◽  
Phil J. Bremer ◽  
Kyoung-Woong Kim ◽  
A. James McQuillan
Keyword(s):  
Ir Spectroscopy ◽  
Metal Ion ◽  
Ion Binding ◽  
Metal Ion Binding

scholarly journals FT-IR Spectroscopy for the Identification of Binding Sites and Measurements of the Binding Interactions of Important Metal Ions with Bovine Serum Albumin

2019 ◽  
Vol 87 (1) ◽  
pp. 5 ◽  
Author(s):  
Hassan Alhazmi
Keyword(s):  
Bovine Serum Albumin ◽  
Ir Spectroscopy ◽  
Serum Albumin ◽  
Metal Ions ◽  
Bovine Serum ◽  
Metal Ion ◽  
Binding Interaction ◽  
Binding Interactions ◽  
Ft Ir ◽  
Ft Ir Spectroscopy

Proteins play crucial roles in the transportation and distribution of therapeutic substances, including metal ions in living systems. Some metal ions can strongly associate, while others show low affinity towards proteins. Consequently, in the present work, the binding behaviors of Ca2+, Ba2+, Ag+, Ru3+, Cu2+ and Co2+ with bovine serum albumin (BSA) were screened. BSA and the metal ions were allowed to interact at physiological pH and their binding interactions were screened by using FT-IR spectroscopy. Spectra were collected by using hydrated films over a range of 4000–400 cm−1. The interaction was demonstrated by a significant reduction in the spectral intensities of the amide I (C=O stretching) and amide II bands (C–N stretching coupled to NH bending) of the protein after complexation with metal ions. The binding interaction was further revealed by spectral shifting of the amide I band from 1651 cm−1 (free BSA) to 1653, 1654, 1649, 1655, 1655, and 1654 cm−1 for BSA–Ca2+, BSA–Ba2+, BSA–Ag+, BSA–Ru3+, BSA–Cu2+ and BSA–Co2+ complexes, respectively. The shifting of the amide I band was due to the interactions of metal ions with the O and N atoms of the ligand protein. Estimation of the secondary protein structure showed alteration in the protein conformation, characterized by a marked decrease (12.9–40.3%) in the α-helix accompanied by increased β-sheet and β-turn after interaction with the metal ions. The interaction results of this study were comparable with those reported in our previous investigation of metal ion–BSA interactions using affinity capillary electrophoresis (ACE), which has proven the accuracy of the FT-IR technique in the measurement of interactions between proteins and metal ions.

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