pH-dependent protein conformational changes in albumin : gold nanoparticle bioconjugates: A spectroscopic study

The conformational changes of bovine serum albumin (BSA) in the albumin:gold nanoparticle bioconjugates were investigated in detail by various spectroscopic techniques including UV-vis absorption, fluorescence, circular dichroism, and Fourier transform infrared spectroscopies. Our studies suggested that albumin in the bioconjugates that was prepared by the common adsorption method underwent substantial conformational changes at both secondary and tertiary structure levels. BSA was found to adopt a more flexible conformational state on the boundary surface of gold nanoparticles as a result of the conformational changes in the bioconjugates. The conformational changes at pH 3.8, 7.0, and 9.0, which corresponded to different isomeric forms of albumin, were investigated, respectively, to probe the pH effect on the conformational changes of BSA in the bioconjugates. The results showed that the pH of the medium influenced the changes greatly and that fluorescence and circular dichroism studies further indicated that the changes were larger at higher pH.

In situ circular dichroic electrochemical study of bilirubin and bovine serum albumin complex

The electrooxidation of bilirubin (BR) and bovine serum albumin (BSA) complexes was studied by in situ circular dichroism (CD) spectroelectrochemistry. The result showed that the mechanism of the whole electrooxidation process of this complex corresponded to electrochemical processes (EE mechanism) in aqueous solution. Some parameters of the process were obtained by double logarithm method, differential method and nonlinear regression method. In visible region, CD spectra of the two enantiomeric components of the complex and their fraction distribution against applied potentials were obtained by singular value decomposition least-square (SVDLS) method. Meanwhile, the distribution of the five components of secondary structure was also obtained by the same method in far-UV region. The peak potential gotten from EE mechanism corresponds to a turning point for the component transition, beyond which the whole reaction reaches a new equilibrium. Under applied positive potentials, the enantiomeric equilibrium between M and P form is broken and M form transfers to its enantiomer of P, while the fraction of alpha-helix increases and that improves the transition to P form.

In vitro study on the binding of neutral red to bovine serum albumin by molecular spectroscopy

In this paper, the binding of neutral red (NR) to bovine serum albumin (BSA) under physiological conditions has been studied by spectroscopy method including fluorescence, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy. The Stern-Volmer fluorescence quenching constant (K-SV), binding constant (K-b) and the number of binding sites (It) were measured by fluorescence quenching method. Fluorescence experiments were also performed at different ionic strengths. It was found K-SV was ionic strength dependent, which indicated the electrostatic interactions were part of the binding forces. The distance r between donor (BSA) and acceptor (NR) was obtained according to Foster's non-radiative energy transfer theory. CD spectroscopy and FT-IR spectroscopy were used to investigate the structural information of BSA molecules on the binding of NR, and the results showed no change of BSA conformation in our experimental conditions.

Detection of anti-human serum albumin antibody using surface plasmon resonance biosensor

The biosensor based on surface plasmon resonance(SPR) technology is a very useful tool to study the interaction between biomolecles. The main advantages of this technique is to "visualize" macromolecular interactions directly in real time, and in a label-free mode rather than indirect methods like enzyme-linked immunosorbent assays (ELISAs). We immobilize human serum albumin (HSA) to the carboxymethyldextran-modified sensor chip surface covalently to detect the activity of anti-HSA in serum, and regenerate the surface with .1 mol/L phosphoric acid. The results show that SPR biosensor can detect the activity of anti-HSA in real-time quickly and the sensor chip can be used over 100 cycles.

Studies of Gd-diethylenetriamine pentaacetic acid in boving serum albumin solution by nuclear magnetic resonance spectroscopy

In this paper, the water relaxation enhancement behavior of Gd-diethylenetriamine pentaacetic acid(DTPA) in water and in aqueous solution of bovine serum albumine(BSA) has been studied. The T-1 relaxivity of Gd-DTPA in BSA solution is higher than that in aqueous solution. The results indicate that Gd-DTPA can integrate non-covalently with BSA mainly in forms of (Gd-DTPA) . BSA, (Gd-DTPA)(2) . BSA, for which the apparent equilibrium constant is 0.026 mmol(-1).L,0.0018 mmol(-2).L-2 respectively. This method would be used to study the interactivities between protein and contrast agent.

Electric field-induced conformational transition of bovine serum albumin from alpha-helix to beta-sheet

The irreversible conformational transition of bovine serum albumin (BSA) from alpha-helix to beta-sheet, induced by electric field near the electrode surface, was monitored by circular dichroism (CD) with a long optical path thin layer cell (LOPTLC).

The electrochemically induced conformational transition of disulfides in bovine serum albumin studied by thin layer circular dichroism spectroelectrochemistry

The conformational transition of disulfides in bovine serum albumin (BSA) induced by electrochemical redox reaction of disulfides were monitored by in-situ circular dichroism (CD) spectroelectrochemistry, with a long optical path thin layer cell and analyzed by a singular value decomposition least square (SVDLS) method. Electrochemical reduction of disulfides drives the left-handed conformation of disulfides changed into the right-handed. At open circuit, eight of the 17 disulfides were of left-handed conformation. Four of the 17 disulfides took part in the electrochemical reduction with an EC mechanism. Only one-fourth of the reduced disulfides returned to left-handed conformation in the re-oxidation process. Some parameters of the electrochemical reduction process, i.e. the number of electrons transferred and electron transfer coefficient, n=8, alphan=0.15, apparent formal potential, E-1(0') = -0.65(+/-0.01) V, standard heterogeneous electron transfer rate constant, k(1)(0) = (2.84 +/- 0.14)x 10(-5) cm s(-1) and chemical reaction equilibrium constant, K-c=(5.13 +/- 0.12) x 10(-2), were also obtained by double logarithmic analysis based on the near-UV absorption spectra with applied potentials. (C) 2001 Elsevier Science B.V. All rights reserved.

NMR relaxation studies of GdDTPA in human serum albumin solution

The water relaxation enhancement behavior of GdDTPA in human serum albumin (HSA) solution has been studied. The results indicate that GdDTPA can integrate noncovalently with HSA, mainly in forms of (GdDTPA)HSA and (GdDTPA),HSA, for which the apparent equilibrium constants are 0.05 mM(-1) and 0.02 mM(-2), respectively. (C) 1999 Elsevier Science Ltd. All rights reserved.

Studies on the interaction of rare earth ions with BSA

Studies on the bounding character of rare earth ions with borine serum albumin(BSA) are significant for understanding the state of rare earth ions in body and their effects on the structure and function of protein. The fluorescence spectrum and pH potentiometry showed consistent results of apparent complexion constant of Tb-2 . BSA. The equilibrium dialysis showed that there are two specific binding sites and more than six non-specific binding sites of RE ions onto BSA molecule with the conditional stable constants lg K-1 = 5. 157 and lgK(2) = 3. 435. Na-23 NMR studies revealed that the BSA peptide chain bound to RE ions was expanded and the mobility of its molecular backbone was increased.

NMR studies of Gd(DTPA-BIN) in human serum albumin solution

Stable gadolinium complexes, such as Gd(DTPA) and Gd(DOTA), are usually used as the contrast agents for magnetic resonance imaging(MRI). Reported here are the enhanced relaxation properties of a novel gadolinium complex, diethylene-triaminopentaacetate Lis (isoniazid) [Gd(DTPA-BIN)], in aqueous and in human serum albumin(HSA) solution, which indicates that (1) two Gd(DTPA-BIN) can integrate non-covalently with one HSA with an equilibrium constant of 0. 02 mmol(-2) . L-2 ; (2) the relaxivities are 3. 28 and 4. 92 mmol(-1) . L . s(-1) for the free Gd(DTPA-BIN) and the [Gd(DTPA-BIN)](2), HSA conjugator, respectively; (3) the rotational correlation time of protein conjugator is notably higher than that of the free complex, The above results may imply that Gd(DTPA-BIN) has a higher tissue selectivity than that of its parent Gd(DTPA).

Fluorescence spectroscopy studies on the action of rare earths with bovine serum albumin in multimolecular system

In this paper. the interaction of tripositive terbium ions (Tb3+) with bovine serum albumin (BSA) has been investigated in the presence of some alkaline earth metals and citric acid (Cit) by using fluorescence spectroscopy. The results show that Tb-31. BSA and Cit can form ternary complex BSA . Tb-2. Cit(4) in mu =0.1 mol/l NaCl. pH6.3 hexamethylenetetramine buffer. Other tare earths are able to compete for the same binding site in BSA with Tb3+-. The sequence of con,petition is Eu3+>Pr3+>Yb3+>Gd3+>La3+>Ca2+ and Mg2+ cannot replace Tb bound to BSA.

IN-SITU UV-VISIBLE SPECTROELECTROCHEMICAL AND CIRCULAR DICHROIC ELECTROCHEMICAL STUDY OF BILIRUBIN AND BILIRUBIN PLUS HUMAN SERUM-ALBUMIN COMPLEX

The electro-oxidation of bilirubin (BR) in aqueous solution was investigated by cyclic voltammetry and in-situ thin-layer spectroelectrochemical techniques, It was found that both oxidation processes of BR are two electron transfer reactions. A mechanism