Characterization of the mangiferin–human serum albumin complex by spectroscopic and molecular modeling approaches

doi:10.1016/j.jpba.2008.12.017

Journal of Pharmaceutical and Biomedical Analysis

Volume 49, Issue 3, 5 April 2009, Pages 753-759

https://doi.org/10.1016/j.jpba.2008.12.017 Get rights and content

Abstract

The interactions between mangiferin and human serum albumin (HSA) were investigated by spectroscopy and molecular modeling. The results proved the formation of complex between mangiferin and HSA. Hydrophobic interaction dominated in the association reaction. Mangiferin statically quenched the fluorescence of HSA in a concentration dependent manner positively deviating from the linear Scatchard equation. The binding of mangiferin to HSA lead to changes in the conformation of HSA according to synchronous fluorescence spectra, FT-IR, UV–vis and CD data. The presence of amino acids and metal ion affected the binding constant of mangiferin–HSA complex. Computational mapping of the possible binding sites of mangiferin revealed the molecule to be bound in the large hydrophobic cavity of subdomain IIA.

Introduction

Mangiferin (Fig. 1), a xanthone glucoside, occurs widely in the bark of Mangifera indica (Family: Anacardiaceae, Genus: Mangifera) [1]. Mangiferin is recommended in the Indian systems of medicine [2] for the treatment of immunodeficiency diseases such as arthritis, hepatitis, cardiac, mental disorders, cancer, autoimmune disorders, arteriosclerosis and coronary heart disease [3]. Some reports have showed that mangiferin possesses antioxidant [4], antitumor [5], antiviral [6] and immunomodulatory activities [7]. Furthermore, mangiferin has recently been shown to have antidiabetic activity in KK/Ay mice, a genetic model of non-insulin-dependent diabetes mellitus (NIDDM) with hyperinsulinemia [8], [9]. Understanding the interaction between mangiferin and protein is of major pharmaceutical and clinical importance. Investigating the interaction of mangiferin to protein can also elucidate the properties of drug-protein complex.

Human serum albumin (HSA) is found to be major protein components of blood plasma [10]. HSA constitutes over half of the total plasma proteins, a concentration of 35–50 g/L, in a healthy individual [11]. It is a globular protein consisting of a single peptide chain of 585 amino acids and is considered to have three specific binding sites (I–III) for high affinity binding of drugs. Each of the sites consists of two subdomains (A and B), and is stabilized by 17 disulfide bridges. The primary pharmacokinetics function of HSA is participating in absorption, distribution, metabolism and excretion of drugs. Therefore, it is often used as a probe to investigate the binding properties of drugs with HSA. It has been shown that the distribution, free concentration and the metabolism of various drugs can be significantly altered as a result of their binding to HSA [12]. Therefore, investigating the interaction of drugs and serum albumins was significant for knowing the transport and distribution of drugs in body, and for clarifying the action mechanism and pharmaceutical dynamics. Yet, no work has been published for the mechanism of the interactions and detailed physicochemical characterizations of mangiferin binding to HSA.

In this study, the interaction of mangiferin with human serum albumin was investigated by biophysical methods mainly fluorescence, UV–vis, FT-IR and CD studies, serve as aids to better understand the mechanism of the drug binding to HSA. The results have been discussed on the binding parameters, the identification of binding sites, the effect of mangiferin on the conformation of HSA, and the nature of forces involved in the interactions. Furthermore, the binding site of mangiferin to HSA was also discussed using automated molecular docking approach.

Section snippets

Materials

HSA and mangiferin were purchased from Sigma. HSA was essentially fatty acid free, and its molecular weight was assumed to be 66,500 to calculate the molar concentrations. All HSA solutions were prepared in a pH 7.40 buffer solution, and the HSA stock solution was kept in the dark at 4 °C. All reagents were of analytical reagent grade. NaCl (1.0 M) solution was used to maintain the ionic strength at 0.1. The buffer (pH 7.40) consisted of Tris (0.2 M) and HCl (0.1 M). The pH was measured by using a

Analysis of fluorescence quenching of HSA by mangiferin

Fluorescence quenching is the decrease of the quantum yield of fluorescence from a fluorophore induced by a variety of molecular interactions with quencher molecule. It has been reported that the binding of small molecules to HSA could induce the conformational change of HSA, because the intramolecular forces involved to maintain the secondary structure could be altered, which results in the conformational change of protein [17]. The conformational changes of HSA were evaluated by the

Conclusions

The interactions between mangiferin and HSA were investigated by different optical techniques and molecular modeling in this paper. Results showed that mangiferin quenched the fluorescence of HSA through static quenching mechanism. Hydrophobic interactions played a role in the binding process of mangiferin to HSA. The results of these methods were consonant with each other. Additionally, docking calculations found mangiferin to be located in site I of HSA within subdomain IIA. The biological

References (39)

J.M. Leiro et al.
Biochem. Pharmacol.
(2003)
C. Yoosook et al.
Phytomedicine
(2000)
D.C. Carter et al.
Adv. Protein Chem.
(1994)
I. Petitpas et al.
J. Biol. Chem.
(2001)
C.Q. Jiang et al.
Spectrochim. Acta A
(2003)
W. He et al.
Bioorg. Med. Chem.
(2005)
M.H. Rahman et al.
Biochem. Pharmacol.
(1993)
J.H. Tang et al.
Bioorg. Med. Chem.
(2006)
Y.J. Hu et al.
J. Pharm. Biomed. Anal.
(2004)
F.L. Cui et al.
Bioorg. Med. Chem.
(2004)

Y.Y. Yue et al.

J. Photochem. Photobiol. B

(2008)

X.F. Liu et al.

J. Inorg. Biochem.

(2005)

Y.Q. Wang

J. Lumin.

(2007)

R.N. Chopra et al.

Glossary of Indian Medicinal Plants

(1956)

S. Ghosal et al.

Ind. J. Chem.

(1996)

T. Sato et al.

Chem. Pharm. Bull.

(1992)

S. Guha et al.

Chemotherapy

(1996)

H. Ichiki et al.

Biol. Pharm. Bull.

(1998)

T. Miura et al.

Biol. Pharm. Bull.

(2001)

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Characterization of the mangiferin–human serum albumin complex by spectroscopic and molecular modeling approaches

Abstract

Introduction

Section snippets

Materials

Analysis of fluorescence quenching of HSA by mangiferin

Conclusions

Biochem. Pharmacol.

Phytomedicine

Adv. Protein Chem.

J. Biol. Chem.

Spectrochim. Acta A

Bioorg. Med. Chem.

Biochem. Pharmacol.

Bioorg. Med. Chem.

J. Pharm. Biomed. Anal.

Bioorg. Med. Chem.

J. Photochem. Photobiol. B

J. Inorg. Biochem.

J. Lumin.

Glossary of Indian Medicinal Plants

Ind. J. Chem.

Chem. Pharm. Bull.

Chemotherapy

Biol. Pharm. Bull.

Biol. Pharm. Bull.