Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Jun;49(2):235-255.
doi: 10.1007/s10867-023-09629-z. Epub 2023 Mar 13.

Probing the binding of morin with alpha-2-macroglobulin using multi-spectroscopic and molecular docking approach : Interaction of morin with α2M

Sana Ansari  1 , Mohammad Khalid Zia  1 , Shamila Fatima  1 , Haseeb Ahsan  2 , Fahim H Khan  3
Affiliations

Probing the binding of morin with alpha-2-macroglobulin using multi-spectroscopic and molecular docking approach : Interaction of morin with α2M

Sana Ansari et al. J Biol Phys. 2023 Jun.

Abstract

Alpha-2-macroglobulin (α2M) is an essential antiproteinase that is widely distributed in human plasma. The present study was aimed at investigating the binding of a potential therapeutic dietary flavonol, morin, with human α2M using a multi-spectroscopic and molecular docking approach. Recently, flavonoid-protein interaction has gained significant attention, because a majority of dietary bioactive components interact with proteins, thereby altering their structure and function. The results of the activity assay exhibited a 48% reduction in the antiproteolytic potential of α2M upon interaction with morin. Fluorescence quenching tests unequivocally confirmed quenching in the fluorescence of α2M in the presence of morin, conforming complex formation and demonstrating that the binding mechanism involves a dynamic mode of interaction. Synchronous fluorescence spectra of α2M with morin showed perturbation in the microenvironment around tryptophan residues. Furthermore, structural changes were observed through CD and FT-IR, showing alterations in the secondary structure of α2M induced by morin. FRET further supports the results of the dynamic mode of quenching. Moderate interaction is shown by binding constant values using Stern-Volmer's fluorescence spectroscopy. Morin binds to α2M at 298 K with a binding constant of 2.7 × 104 M-1, indicating the strength of the association. The α2M-morin system was found to have negative ΔG values, which suggests that the binding process was spontaneous. Molecular docking also reveals the different amino acid residues involved in this binding process, revealing that the binding energy is -8.1 kcal/mol.

Keywords: Alpha-2-macroglobulin; Antiproteinase; Flavonol; Morin; Proteinase.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Effect of increasing concentration of morin (5–20 μM) on antiproteinase activity of α2M (20 μM)
Fig. 2
Fig. 2
Effect of morin on absorption spectra of purified human α2M. Curve a, α2M alone (20 μM); curve b-h, human α2M (20 μM) treated with increasing concentration of morin (2, 4, 6, 8, 10, 15, and 20 μM)
Fig. 3
Fig. 3
Intrinsic fluorescence spectra of native and morin treated α2M. Curve a, α2M alone (20 μM); curve b-h, human α2M (20 μM) treated with increasing concentration of morin (2, 4, 6, 8, 10, 15, and 20 μM); curve i, morin alone (20 μM)
Fig. 4
Fig. 4
Stern–Volmer plots for the quenching of α2M (20 μM) by morin at three different temperatures (298, 303, and 310 K)
Fig. 5
Fig. 5
Modified Stern–Volmer plots for the quenching of α2M (20 μM) by morin at three different temperatures (298, 303, and 310 K)
Fig. 6
Fig. 6
Van’t Hoff plots for the temperature dependence of Kb for calculation of thermodynamic parameters
Fig. 7
Fig. 7
Spectral overlap between the fluorescence emission spectrum of α2M with the absorption spectrum of morin, [morin]/[α2M] = 1:1, T = 298 K
Fig. 8
Fig. 8
Circular dichroism spectra of α2M (20 μM) in the absence and presence of different concentrations of morin (10 and 20 μM) at 298 K
Fig. 9
Fig. 9
A Synchronous fluorescence spectra of α2M-morin system for Δλ = 15 nm. Curve a, α2M alone (20 μM); curve b-h, human α2M (20 μM) treated with increasing concentration of morin (2, 4, 6, 8, 10, 15, and 20 μM). B Synchronous fluorescence spectra of α2M-morin system for Δλ = 60 nm. Curve a, α2M alone (20 μM); curve b-h, human α2M (20 μM) treated with increasing concentration of morin (2, 4, 6, 8, 10, 15, and 20 μM)
Fig. 10
Fig. 10
FTIR spectra of native and morin treated α2M. Purified human α2M (20 μM) was incubated with morin (20 μM) concentration at 298 K
Fig. 11
Fig. 11
A Surface view of human α2M depicting the pocket where morin in its pocket. B Ligplot depicting amino acid residues of human α2M involved in interaction with morin
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Ullah A, Munir S, Badshah SL, Khan N, Ghani L, Poulson BG, Jaremko M. Important flavonoids and their role as a therapeutic agent. Molecules. 2020;25(22):5243. doi: 10.3390/molecules25225243. - DOI - PMC - PubMed
    1. López-Yerena A, Perez M, Vallverdú-Queralt A, Escribano-Ferrer E. Insights into the binding of dietary phenolic compounds to human serum albumin and food-drug interactions. Pharmaceutics. 2020;12(11):1123. doi: 10.3390/pharmaceutics12111123. - DOI - PMC - PubMed
    1. Caselli A, Cirri P, Santi A, Paoli P. Morin: a promising natural drug. Curr Med Chem. 2016;23(8):774–791. doi: 10.2174/0929867323666160106150821. - DOI - PubMed
    1. Choudhury A, Chakraborty I, Banerjee TS, Vana DR, Adapa D. Efficacy of morin as a potential therapeutic phytocomponent: insights into the mechanism of action. Int. J. Med. Res. Health Sci. 2017;6(11):175–194.
    1. Choi YA, Yoon YH, Choi K, Kwon M, Goo SH, Cha JS, Song IS. Enhanced oral bioavailability of morin administered in mixed micelle formulation with PluronicF127 and Tween80 in rats. Biol. Pharm. Bull. 2015;38(2):208–217. doi: 10.1248/bpb.b14-00508. - DOI - PubMed

Publication types

LinkOut - more resources

-