Steady-state and time-resolved fluorescence investigation of 2-pyridone and 3-pyridone in solution and their specific binding to human serum albumin

Osama K. Abou-Zied, O. I K Al-Shihi

Research output: Chapter in Book/Report/Conference proceedingConference contribution

5 Citations (Scopus)

Abstract

2-pyridone (2Py) and 3-pyridone (3Py) were examined in different solvents and their binding to human serum albumin (HSA) was studied using steady-state spectroscopy and time-resolved fluorescence. Solvation of 2Py and 3Py by water was examined in binary mixtures of 1,4-dioxane and water. Analysis of the absorption and fluorescence data reveals the solvation of the hydrogen bonding center in 2Py by one water molecule and in 3Py by three water molecules. A zwitterionic tautomer of 3Py is formed in water and shows distinct absorption peaks from the absorption of the neutral tautomer. Fluorescence of 3Py was observed in polar solvents only, whereas 2Py is fluorescent in polar and nonpolar solvents. The absorption and fluorescence spectra of 2Py in different solvents indicate less solute-solvent interaction in nonpolar solvents. This observation was confirmed by the measured longer fluorescence lifetime of 2Py in cyclohexane compared to that in water. The mechanism of binding of 2Py and 3Py as probe ligands to HSA was investigated by following the intensity change and lifetime of HSA fluorescence after excitation at 280 nm. The presence of 2Py and 3Py causes a reduction in the fluorescence intensity and lifetime of HSA. This observation indicates that subdomain IIA binding site (Sudlow site I) is the host of the probes and the reduction in the fluorescence of HSA is due to energy transfer from the Trp-214 residue to the probe in each case. The distance between Trp-214 and each of the probes was calculated using Förster theory for energy transfer to be 1.99 nm for HSA/2Py and 2.44 nm for HSA/3Py. The shorter distance in the former complex indicates more efficient energy transfer than in the latter. This was confirmed by estimating the quenching rate constant (kq) in each complex. kq was calculated to be 1.44 × 1012 M-1S-1 for HSA/2Py and 3.45 × 1011 M-1 s-1 for HSA/3Py. The calculated distances and the kq values indicate a static quenching mechanism operative in the two complexes. The binding constants were estimated to be K = (3.4 ± 0.4) × 104 M-1 for the HSA/2Py complex and K= (2.3 ± 0.3) × 104 M-1 for the HSA/3Py complex. The number of binding sites of HSA was calculated to be one in both complexes. The latter results, along with the quenching results, indicate that both probes, 2Py and 3Py, bind only in Sudlow site I in subdomain IIA.

Original languageEnglish
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume6867
DOIs
Publication statusPublished - 2008
EventMolecular Probes for Biomedical Applications II - San Jose, CA, United States
Duration: Jan 21 2008Jan 22 2008

Other

OtherMolecular Probes for Biomedical Applications II
CountryUnited States
CitySan Jose, CA
Period1/21/081/22/08

Fingerprint

Fluorescence
Energy transfer
Water
Quenching
Solvation
Binding sites
Molecules
Cyclohexane
Binary mixtures
Rate constants
Hydrogen bonds
Ligands
Spectroscopy

Keywords

  • 2-pyridone
  • 3-pyridone
  • Biological probes
  • Human serum albumin
  • Hydroxypyridines
  • Protein-ligand recognition
  • Resonance energy transfer
  • Tautomerization

ASJC Scopus subject areas

  • Engineering(all)

Cite this

Steady-state and time-resolved fluorescence investigation of 2-pyridone and 3-pyridone in solution and their specific binding to human serum albumin. / Abou-Zied, Osama K.; Al-Shihi, O. I K.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 6867 2008. 68670K.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abou-Zied, OK & Al-Shihi, OIK 2008, Steady-state and time-resolved fluorescence investigation of 2-pyridone and 3-pyridone in solution and their specific binding to human serum albumin. in Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 6867, 68670K, Molecular Probes for Biomedical Applications II, San Jose, CA, United States, 1/21/08. https://doi.org/10.1117/12.762218
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title = "Steady-state and time-resolved fluorescence investigation of 2-pyridone and 3-pyridone in solution and their specific binding to human serum albumin",
abstract = "2-pyridone (2Py) and 3-pyridone (3Py) were examined in different solvents and their binding to human serum albumin (HSA) was studied using steady-state spectroscopy and time-resolved fluorescence. Solvation of 2Py and 3Py by water was examined in binary mixtures of 1,4-dioxane and water. Analysis of the absorption and fluorescence data reveals the solvation of the hydrogen bonding center in 2Py by one water molecule and in 3Py by three water molecules. A zwitterionic tautomer of 3Py is formed in water and shows distinct absorption peaks from the absorption of the neutral tautomer. Fluorescence of 3Py was observed in polar solvents only, whereas 2Py is fluorescent in polar and nonpolar solvents. The absorption and fluorescence spectra of 2Py in different solvents indicate less solute-solvent interaction in nonpolar solvents. This observation was confirmed by the measured longer fluorescence lifetime of 2Py in cyclohexane compared to that in water. The mechanism of binding of 2Py and 3Py as probe ligands to HSA was investigated by following the intensity change and lifetime of HSA fluorescence after excitation at 280 nm. The presence of 2Py and 3Py causes a reduction in the fluorescence intensity and lifetime of HSA. This observation indicates that subdomain IIA binding site (Sudlow site I) is the host of the probes and the reduction in the fluorescence of HSA is due to energy transfer from the Trp-214 residue to the probe in each case. The distance between Trp-214 and each of the probes was calculated using F{\"o}rster theory for energy transfer to be 1.99 nm for HSA/2Py and 2.44 nm for HSA/3Py. The shorter distance in the former complex indicates more efficient energy transfer than in the latter. This was confirmed by estimating the quenching rate constant (kq) in each complex. kq was calculated to be 1.44 × 1012 M-1S-1 for HSA/2Py and 3.45 × 1011 M-1 s-1 for HSA/3Py. The calculated distances and the kq values indicate a static quenching mechanism operative in the two complexes. The binding constants were estimated to be K = (3.4 ± 0.4) × 104 M-1 for the HSA/2Py complex and K= (2.3 ± 0.3) × 104 M-1 for the HSA/3Py complex. The number of binding sites of HSA was calculated to be one in both complexes. The latter results, along with the quenching results, indicate that both probes, 2Py and 3Py, bind only in Sudlow site I in subdomain IIA.",
keywords = "2-pyridone, 3-pyridone, Biological probes, Human serum albumin, Hydroxypyridines, Protein-ligand recognition, Resonance energy transfer, Tautomerization",
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T1 - Steady-state and time-resolved fluorescence investigation of 2-pyridone and 3-pyridone in solution and their specific binding to human serum albumin

AU - Abou-Zied, Osama K.

AU - Al-Shihi, O. I K

PY - 2008

Y1 - 2008

N2 - 2-pyridone (2Py) and 3-pyridone (3Py) were examined in different solvents and their binding to human serum albumin (HSA) was studied using steady-state spectroscopy and time-resolved fluorescence. Solvation of 2Py and 3Py by water was examined in binary mixtures of 1,4-dioxane and water. Analysis of the absorption and fluorescence data reveals the solvation of the hydrogen bonding center in 2Py by one water molecule and in 3Py by three water molecules. A zwitterionic tautomer of 3Py is formed in water and shows distinct absorption peaks from the absorption of the neutral tautomer. Fluorescence of 3Py was observed in polar solvents only, whereas 2Py is fluorescent in polar and nonpolar solvents. The absorption and fluorescence spectra of 2Py in different solvents indicate less solute-solvent interaction in nonpolar solvents. This observation was confirmed by the measured longer fluorescence lifetime of 2Py in cyclohexane compared to that in water. The mechanism of binding of 2Py and 3Py as probe ligands to HSA was investigated by following the intensity change and lifetime of HSA fluorescence after excitation at 280 nm. The presence of 2Py and 3Py causes a reduction in the fluorescence intensity and lifetime of HSA. This observation indicates that subdomain IIA binding site (Sudlow site I) is the host of the probes and the reduction in the fluorescence of HSA is due to energy transfer from the Trp-214 residue to the probe in each case. The distance between Trp-214 and each of the probes was calculated using Förster theory for energy transfer to be 1.99 nm for HSA/2Py and 2.44 nm for HSA/3Py. The shorter distance in the former complex indicates more efficient energy transfer than in the latter. This was confirmed by estimating the quenching rate constant (kq) in each complex. kq was calculated to be 1.44 × 1012 M-1S-1 for HSA/2Py and 3.45 × 1011 M-1 s-1 for HSA/3Py. The calculated distances and the kq values indicate a static quenching mechanism operative in the two complexes. The binding constants were estimated to be K = (3.4 ± 0.4) × 104 M-1 for the HSA/2Py complex and K= (2.3 ± 0.3) × 104 M-1 for the HSA/3Py complex. The number of binding sites of HSA was calculated to be one in both complexes. The latter results, along with the quenching results, indicate that both probes, 2Py and 3Py, bind only in Sudlow site I in subdomain IIA.

AB - 2-pyridone (2Py) and 3-pyridone (3Py) were examined in different solvents and their binding to human serum albumin (HSA) was studied using steady-state spectroscopy and time-resolved fluorescence. Solvation of 2Py and 3Py by water was examined in binary mixtures of 1,4-dioxane and water. Analysis of the absorption and fluorescence data reveals the solvation of the hydrogen bonding center in 2Py by one water molecule and in 3Py by three water molecules. A zwitterionic tautomer of 3Py is formed in water and shows distinct absorption peaks from the absorption of the neutral tautomer. Fluorescence of 3Py was observed in polar solvents only, whereas 2Py is fluorescent in polar and nonpolar solvents. The absorption and fluorescence spectra of 2Py in different solvents indicate less solute-solvent interaction in nonpolar solvents. This observation was confirmed by the measured longer fluorescence lifetime of 2Py in cyclohexane compared to that in water. The mechanism of binding of 2Py and 3Py as probe ligands to HSA was investigated by following the intensity change and lifetime of HSA fluorescence after excitation at 280 nm. The presence of 2Py and 3Py causes a reduction in the fluorescence intensity and lifetime of HSA. This observation indicates that subdomain IIA binding site (Sudlow site I) is the host of the probes and the reduction in the fluorescence of HSA is due to energy transfer from the Trp-214 residue to the probe in each case. The distance between Trp-214 and each of the probes was calculated using Förster theory for energy transfer to be 1.99 nm for HSA/2Py and 2.44 nm for HSA/3Py. The shorter distance in the former complex indicates more efficient energy transfer than in the latter. This was confirmed by estimating the quenching rate constant (kq) in each complex. kq was calculated to be 1.44 × 1012 M-1S-1 for HSA/2Py and 3.45 × 1011 M-1 s-1 for HSA/3Py. The calculated distances and the kq values indicate a static quenching mechanism operative in the two complexes. The binding constants were estimated to be K = (3.4 ± 0.4) × 104 M-1 for the HSA/2Py complex and K= (2.3 ± 0.3) × 104 M-1 for the HSA/3Py complex. The number of binding sites of HSA was calculated to be one in both complexes. The latter results, along with the quenching results, indicate that both probes, 2Py and 3Py, bind only in Sudlow site I in subdomain IIA.

KW - 2-pyridone

KW - 3-pyridone

KW - Biological probes

KW - Human serum albumin

KW - Hydroxypyridines

KW - Protein-ligand recognition

KW - Resonance energy transfer

KW - Tautomerization

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U2 - 10.1117/12.762218

DO - 10.1117/12.762218

M3 - Conference contribution

AN - SCOPUS:42249111888

SN - 9780819470423

VL - 6867

BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE

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