Study on the interactions of kaempferol and quercetin with intravenous immunoglobulin by fluorescence quenching, fourier transformation infrared spectroscopy and circular dichroism spectroscopy.
The interactions of kaempferol and quercetin with intravenous immunoglobulin (IVIG) were studied in vitro by spectroscopic methods including fluorescence spectra, Fourier transformation infrared (FT-IR) spectra and circular dichroism (CD) spectra. The binding parameters for the reactions calculated according to the Sips equation suggested that the bindings of IVIG to kaempferol and quercetin were characterized by two binding sites with the average affinity constants K(o) at 1.032 x 10(4) M(-1) and 1.849 x 10(4) M(-1), respectively. The binding of IVIG with quercetin is stronger than that of IVIG with kaempferol. They were of non-specific and weak drug-protein interactions. Docking was used to calculate the interaction modes between kaempferol and quercetin with IVIG. The secondary structural compositions of free IVIG and its kaempferol, quercetin complexes were calculated by the FT-IR difference spectra, self-deconvolution, second derivative resolution enhancement and the curve-fitting procedures of amide I band respectively, which are in good agreement with the analyses of CD spectra. The effect of 3'-OH substituent in quercetin is distinct between the interactions of IVIG with kaempferol and quercetin for the secondary structure of the protein. The observed spectral changes indicate a partial unfolding of the protein structure, but the typical beta structural conformation of IVIG is still retentive in the presence of both drugs in aqueous solution. The average binding distances between the chromophores of IVIG with kaempferol (4.30 nm) and quercetin (4.35 nm) were obtained on the basis of the theory of Förster energy transfer. IVIG can serve as transport protein (carrier) for kaempferol and quercetin.
Liu
Yong-Chun
YC
College of Chemistry and Chemical Engineering, State Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou, PR China.
Yang
Zheng-Yin
ZY
Du
Juan
J
Yao
Xiao-Jun
XJ
Lei
Rui-Xia
RX
Zheng
Xu-Dong
XD
Liu
Jian-Ning
JN
Hu
Huai-Sheng
HS
Li
Hong
H
D002942:Circular Dichroism; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D016756:Immunoglobulins, Intravenous / Q000737:chemistry*; D044949:Kaempferols / Q000737:chemistry*; D008958:Models, Molecular; D008968:Molecular Conformation; D011794:Quercetin / Q000737:chemistry*; D012709:Serum Albumin / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared
D007074:Immunoglobulin G; D016756:Immunoglobulins, Intravenous; D044949:Kaempferols; D012709:Serum Albumin; C006552:kaempferol; D011794:Quercetin