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Binding of Two Tetrasulfophthalocyanines (Fe(III) and Metal-Free) to Lysozyme: Fluorescence Spectroscopic and Computational Approach

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Abstract

The interactions between tetrasulfophthalocyanines and lysozyme were studied using fluorescence spectroscopic and computational analyses. Lysozyme has been found to be widely studied as an anticancer agent, however, there are few reports of its interaction with phthalocyanines. Fe(III) tetrasulfophthalocyanine (FeTSPc) and free base tetrasulfophthalocyanine (TSPc) used in this study, were synthesized by our research group. Experimental results suggested that the metalled complex FeTSPc has a much higher affinity than TSPc. The binding stoichiometry between each tetrasulfophthalocyanine and lysozyme was 1:1. Stern−Volmer analysis suggested that the fluorescence quenching proceedes through a static process. Binding thermodynamics (ΔG, ΔH and ΔS) confirmed that mainly hydrogen bonds, van der Waals, and electrostatic forces are responsible for the binding process. We carried out molecular dynamics simulations, molecular docking, and binding energy calculations. Molecular dynamics simulations yielded the most populated cluster of lysozyme structures, and a representative structure from this cluster was used for the docking studies with these phthalocyanines. 1000 poses were generated for each ligand. The strudtures of the resulting complexes revealed that Arg 73 and Arg 112 are important for the binding affinity of the tetrasulfophthalocyanines, generating mainly an electrostatic favorable environment for the SO3− groups. In addition, hydrophobic contacts were involved with Trp 62, Trp 63 and Trp 108, explaining the fluorescence quenching observed experimentally. Binding energies were determined for these models, confirming that the interactions with lysozyme were more favorable for FeTSPc compared to TSPc. The understanding of the molecular mechanisms is relevant to characterize the nature of tetrasulfophthalocyanines in photodynamic therapy.

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Abbreviations

A(280 nm, 1%) :

masss extinction coefficient

APBS:

Adaptive Poisson-Boltzmann Solver

ΔGCoul :

Coulombic energy

ΔGelec :

electrostatic energy

ΔGcalc :

binding free energy

ΔG:

Gibbs free energy change

ΔGnon-elec :

non-electrostatic energy

ΔGsolv :

solvation energy

ΔH:

enthalpy change

ΔS:

entropy change

ΔSASA:

solvent-accessible surface area change

FeTSPc:

Fe(III) tetrasulfophthalocyanine

F0 or F:

fluorescence intensities

γ:

interfacial tension coefficient

Kb :

binding equilibrium constant

KSV :

Stern-Volmer constant

kq :

quenching rate constant

λexc :

excitation wavelength

λem :

emission wavelength

NAMD:

Not Another Molecular Dynamics Program

n :

number of binding sites

Q:

molar concentration

R:

gas constant

RMSD:

Root-Mean-Square Deviation

T:

absolute temperature

τ0 :

average lifetime of biomolecules

TSPc:

free base tetrasulfophthalocyanine

UV-Vis:

Ultraviolet-Visible

VMD:

Visual Molecular Dynamics

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Acknowledgments

The authors thank Dr. Nina Pastor Colón, for helpful discussions and advise as well as her help with the revision of the English language. J.O.V.E thanks CONACyT for the posgraduate scholarship No. 130442 for the studies of Master of Sciences (Chemistry). The Laboratorio de Visualización y Cómputo Paralelo at Universidad Autónoma Metropolitana Iztapalapa is acknowledged for computing time.

Availability of Data and Material

The authors have no any objection for availability of data and materials.

Code Availability

Hen egg-white Lysozyme was purchased from Sigma Chemical Co. (lot. 73H7045).

Crystal structure of Hen egg-white lysozyme from Gallus Gallus. https://www.rcsb.org/structure/2VB1

ACD/ChemSketch Freeware http://www.acdlabs.com/resources/freeware/chemsketch/

Visual Molecular Dynamics (VMD) https://www.ks.uiuc.edu/Research/vmd/

NAMD https://www.ks.uiuc.edu/Research/namd/

Autodock Vina http://vina.scripps.edu

Maestro https://www.schrodinger.com/products/maestro

Charmm-gui web server http://www.charmm-gui.org

PDB2PQR http://server.poissonboltzmann.org/pdb2pqr

APBS http://www.poissonboltzmann.org/

Funding

This research was supported by Universidad Autónoma Metropolitana Iztapalapa.

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Authors and Affiliations

  1. Departamento de Química, Universidad Autónoma Metropolitana-Iztapalapa, Avenida San Rafael Atlixco 186, Colonia Vicentina, 09340 Iztapalapa, Ciudad de México, México

    Jonathan Osiris Vicente-Escobar, Miguel Ángel García-Sánchez, Iris N. Serratos & Salvador Ramón Tello-Solís

  2. Facultad de Farmacia, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, México

    César Millán-Pacheco

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  1. Jonathan Osiris Vicente-Escobar
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  2. Miguel Ángel García-Sánchez
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  4. César Millán-Pacheco
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  5. Salvador Ramón Tello-Solís
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Contributions

Jonathan Osiris Vicente-Escobar: investigation, analysis, writing; Miguel Ángel García-Sánchez: investigation, review, formal analysis, resources; Iris N. Serratos: conceptualization, visualization, investigation, software, formal analysis, writing, review & editing, César Millán-Pacheco: investigation, software, formal analysis; Salvador R. Tello-Solís: conceptualization, project administration, visualization, methodology, investigation, formal analysis, writing, review, editing, & resources.

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Correspondence to Salvador Ramón Tello-Solís.

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Vicente-Escobar, J.O., García-Sánchez, M.Á., Serratos, I.N. et al. Binding of Two Tetrasulfophthalocyanines (Fe(III) and Metal-Free) to Lysozyme: Fluorescence Spectroscopic and Computational Approach. J Fluoresc 31, 787–796 (2021). https://doi.org/10.1007/s10895-021-02710-7

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