Effect of Bovine Serum Albumin on the Structure of DSPE Monolayer†

doi:10.7503/cjcu20170359

Abstract

Abstract:

The effect of bovine serum albumin(BSA) on the 1,2-distearoyl-sn-glycero-3-phosphoethanolamine(DSPE) monolayer at air-water interface was investigated by Langmuir-blodgett(LB) technique and atomic force microscopy(AFM) observation. The surface pressure-mean molecular area(π-A) isotherms, adsorption curves and compression-expansion cycle curves of DSPE monolayers were obtained by changing the pH values and the concentration of BSA. The results indicated that the presence of BSA in the subphase had a great influence on the compressibility, stability and phase transition behavior of DSPE monolayers. The adsorption kinetic indicated that BSA was adsorbed onto DSPE monolayers and a threshold value existed, which was affected by the pH value. Analysis of experimental data showed that most hydrophobic residues of BSA were exposed to water at pH=3, so the interaction between BSA and DSPE was strongest. However, the interaction was the weakest at pH=7 because less hydrophobic residues were exposed to water. The change of surface morphology structure observed by AFM was consistent with the results obtained from curves. This study provides an important experimental basis and theoretical basis for understanding the interaction mechanism between serum proteins and phospholipid molecules.

Key words: Bovine serum albumin(BSA), 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine(DSPE), Surface pressure, Interaction

CLC Number:

O641
O621

TrendMD:

XU Guoqing, HAO Changchun, HE Jianzhen, ZHANG Lei, SUN Runguang. Effect of Bovine Serum Albumin on the Structure of DSPE Monolayer^†[J]. Chem. J. Chinese Universities, 2017, 38(12): 2238.

Figures/Tables 12

Fig.1 Molecular structure of DSPE

Fig.2 Surface pressure(π)-mean molecular area(A) isotherms of DSPE monolayers (A) pH=3; (B) pH=5; (C) pH=7; (D) pH=9.

Fig.3 Mean molecular area(ΔA)-BSA concentration[c(BSA)] isotherms of DSPE monolayers(A) 20 mN/m; (B) 40 mN/m.

Fig.4 Surface pressure(π)-time(t) curves of pure DSPE monolayers and DSPE containing BSA(20, 50 nmol/L) monolayers on PBS subphase(A) pH=3; (B) pH=5; (C) pH=7; (D) pH=9.

Table 1 Parameter of π-t curves for pure DSPE monolayers and DSPE containing BSA monolayers at different subphase pH values*

pH	π/(mN·m^-1)
pH	π₀	π₁	π₂	Δπ₁	Δπ₂
3	12.550	18.953	24.804	6.403	12.254
5	14.975	18.704	22.717	3.729	7.742
7	14.842	16.426	17.711	1.584	2.869
9	15.551	18.230	18.516	2.679	2.965

Fig.5 Adsorption kinetics of BSA at different subphase pH valuesc(BSA)=50 nmol/L. Curved lines are the fitting curves using eq.(1). Zero time is adjusted to the time when the rising of pressure starts.

Table 2 Fitting parameters using eq.(1) of adsorption kinetics of BSA at different subphase pH values*

pH	10⁴ k/s^-1	a/(mN·m^-1)	b/(mN·m^-1)	R²
3	8.967×10	17.494	25.148	0.998
5	3.598×10	17.546	23.169	0.998
7	4.797×10	16.322	18.372	0.997
9	3.943×10	17.911	18.667	0.953

Fig.6 Stability of pure DSPE monolayers and DSPE containing BSA(20 nmol/L) monolayers at different subphase pH values(A) pH=3; (B) pH=5; (C) pH=7; (D) pH=9.

Fig.7 Compression-expansion cycles of pure DSPE and DSPE containing BSA at different subphase pH values(A) pH=3; (B) pH=5; (C) pH=7; (D) pH=9.

Fig.8 Model of the process of successive compression-decompression cycles

Fig.9 Compressibility coefficient(Cs-1)-surface pressure(π) curves of DSPE monolayers (A) pH=3; (B) pH=5; (C) pH=7; (D) pH=9.

Fig.10 AFM images of pure DSPE and mixed DSPE/BSA monolayers at different π and pH values(A) DSPE, pH=3, π=20 mN/m; (B) DSPE, pH=3, π=40 mN/m; (C) DSPE-BSA, pH=3, π=20 mN/m; (D) DSPE-BSA, pH=3, π=40 mN/m; (E) DSPE, pH=5, π=20 mN/m; (F) DSPE, pH=5, π=40 mN/m; (G) DSPE-BSA, pH=5, π=20 mN/m; (H) DSPE-BSA, pH=5, π=40 mN/m; (I) DSPE, pH=7, π=20 mN/m; (J) DSPE, pH=7, π=40 mN/m; (K) DSPE-BSA, pH=7, π=20 mN/m; (L) DSPE-BSA, pH=7, π=40 mN/m; (M) DSPE, pH=9, π=20 mN/m; (N) DSPE, pH=9, π=40 mN/m; (O) DSPE-BSA, pH=9, π=20 mN/m; (P) DSPE-BSA, pH=9, π=40 mN/m. c(BSA)=20 nmol/L. Scanning range: 15 μm×15 μm

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