Institute of Food Research, Norwich Research Park, Colney Lane, Norwich NR4 7UA, UK.
Langmuir. 2010 Jun 15;26(12):9782-93. doi: 10.1021/la1000446.
It is increasingly recognized that changes in the composition of the oil-water interface can markedly affect pancreatic lipase adsorption and function. To understand interfacial mechanisms determining lipase activity, we investigated the adsorption behavior of bile salts and pancreatic colipase and lipase onto digalactosyldiacylglycerol (DGDG) and dipalmitoylphosphatidylcholine (DPPC) monolayers at the air-water interface. The results from Langmuir trough and pendant drop experiments showed that a DGDG interface was more resistant to the adsorption of bile salts, colipase, and lipase compared to that of DPPC. Atomic force microscopy (AFM) images showed that the adsorption of bile salts into a DPPC monolayer decreased the size of the liquid condensed (LC) domains while there was no visible topographical change for DGDG systems. The results also showed that colipase and lipase adsorbed exclusively onto the mixed DPPC-bile salt regions and not the DPPC condensed phase. When the colipase and lipase were in excess, they fully covered the mixed DPPC-bile salt regions. However, the colipase and lipase coverage on the mixed DGDG-bile salt monolayer was incomplete and discontinuous. It was postulated that bile salts adsorbed into the DPPC monolayers filling the gaps between the lipid headgroups and spacing out the lipid molecules, making the lipid hydrocarbon tails more exposed to the surface. This created hydrophobic patches suitable for the binding of colipase and lipase. In contrast, bile salts adsorbed less easily into the DGDG monolayer because DGDG has a larger headgroup, which has strong intermolecular interactions and the ability to adopt different orientations at the interface. Thus, there are fewer hydrophobic patches that are of sufficient size to accommodate the colipase on the mixed DGDG-bile salt monolayer compared to the mixed DPPC-bile salt regions. The results from this work have reinforced the hypothesis that the interfacial molecular packing of lipids at the oil-water interface influences the adsorption of bile salts, colipase, and lipase, which in turn impacts the rate of lipolysis.
人们越来越认识到,油水界面组成的变化可以显著影响胰脂肪酶的吸附和功能。为了了解决定脂肪酶活性的界面机制,我们研究了胆汁盐和胰腺辅脂酶以及脂肪酶在二半乳糖基二酰基甘油(DGDG)和二棕榈酰基磷脂酰胆碱(DPPC)单层在气液界面上的吸附行为。Langmuir 槽和悬滴实验的结果表明,与 DPPC 相比,DGDG 界面更能抵抗胆汁盐、辅脂酶和脂肪酶的吸附。原子力显微镜(AFM)图像显示,胆汁盐在 DPPC 单层中的吸附减小了液体凝聚(LC)区域的尺寸,而对于 DGDG 体系没有可见的形貌变化。结果还表明,辅脂酶和脂肪酶仅吸附在 DPPC-胆汁盐混合区域,而不吸附 DPPC 凝聚相。当辅脂酶和脂肪酶过量时,它们完全覆盖 DPPC-胆汁盐混合区域。然而,辅脂酶和脂肪酶在混合 DGDG-胆汁盐单层上的覆盖是不完整和不连续的。有人推测,胆汁盐吸附到 DPPC 单层中,填补了脂质头部基团之间的空隙,并使脂质分子间隔开来,使脂质烃尾更暴露于表面。这就形成了适合辅脂酶和脂肪酶结合的疏水区。相比之下,胆汁盐不易吸附到 DGDG 单层中,因为 DGDG 的头部基团较大,具有较强的分子间相互作用和在界面上采用不同取向的能力。因此,与 DPPC-胆汁盐混合区域相比,混合 DGDG-胆汁盐单层上适合容纳辅脂酶的疏水区数量较少。这项工作的结果加强了这样一种假设,即在油水界面处脂质的界面分子排列影响胆汁盐、辅脂酶和脂肪酶的吸附,进而影响脂肪分解的速度。