Mardilovich Anastasia, Kokkoli Efrosini
Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Langmuir. 2005 Aug 2;21(16):7468-75. doi: 10.1021/la0468085.
Planar-supported lipid bilayers have attracted enormous attention because of their properties as model cell membranes, which can be employed in a variety of fundamental biological studies and medical devices. Furthermore, the development of patterned biological interfaces is of great practical and scientific interest because of their potential applications in the field of biosensors, drug screening, tissue engineering, and medical implants. In this study, mica-supported membranes were constructed from biomimetic peptide-amphiphiles and their mixtures with lipidated poly(ethylene glycol) (PEG120) molecules or 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) phospholipids using the Langmuir-Blodgett technique. The two peptide-amphiphiles used in this study were a fibronectin-mimetic with the PHSRN(SG)(3)SGRGDSP headgroup (referred to as PHSRN-GRGDSP) that contains both the primary (GRGDSP) and the synergy (PHSRN) recognition sites for alpha(5)beta(1) integrins and a peptide-amphiphile that mimics a fragment of the N-terminus of the fractalkine receptor (referred to as NTFR). Compression isotherms of the peptide-amphiphiles and their mixtures with PEG120 at the air/water interface were recorded and analyzed to evaluate the extent of miscibility in the two-component LB films. Domain formation in mica-supported bilayers constructed from mixtures of peptide-amphiphiles and lipidated PEG120 or DPPC was observed using atomic force microscopy. In PHSRN-GRGDSP/PEG120 mixtures deposited from an aqueous subphase at pH 7, concentration-dependent phase separation was observed on the AFM images. The NTFR/PEG120 and NTFR/DPPC mixtures deposited at pH 10 exhibited extensive lateral phase separation at all mixture compositions, whereas at deposition pH 7 the concentrations of NTFR/DPPC examined here were well mixed.
平面支撑的脂质双层因其作为模型细胞膜的特性而备受关注,可用于各种基础生物学研究和医疗设备。此外,图案化生物界面的发展具有重大的实际和科学意义,因为它们在生物传感器、药物筛选、组织工程和医疗植入物领域具有潜在应用。在本研究中,使用朗缪尔-布洛杰特技术,由仿生肽两亲物及其与脂化聚乙二醇(PEG120)分子或1,2-二棕榈酰-sn-甘油-3-磷酸胆碱(DPPC)磷脂的混合物构建云母支撑膜。本研究中使用的两种肽两亲物分别是具有PHSRN(SG)(3)SGRGDSP头基的纤连蛋白模拟物(称为PHSRN-GRGDSP),其包含α(5)β(1)整合素的主要(GRGDSP)和协同(PHSRN)识别位点,以及一种模拟趋化因子受体N端片段的肽两亲物(称为NTFR)。记录并分析了肽两亲物及其与PEG120在空气/水界面的压缩等温线,以评估两组分LB膜中的混溶程度。使用原子力显微镜观察了由肽两亲物与脂化PEG120或DPPC的混合物构建的云母支撑双层中的畴形成。在pH 7的水相下层沉积的PHSRN-GRGDSP/PEG120混合物中,在AFM图像上观察到浓度依赖性相分离。在pH 10沉积的NTFR/PEG120和NTFR/DPPC混合物在所有混合物组成下均表现出广泛的横向相分离,而在沉积pH 7时,此处检测的NTFR/DPPC浓度混合良好。