LIPID AND PROTEIN EFFECTS ON MONOLAYER STABILITY

脂质和蛋白质对单层稳定性的影响

• 批准号: 6490551
• 负责人: Joseph Anthony Zasadzinski
• 金额: $ 23.47万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6490551
• 关键词: X ray crystallography; atomic force microscopy; biophysics; biotechnology; blood proteins; chemical structure function; combinatorial chemistry; fluorescence microscopy; intermolecular interaction; lipid solubility; lipid structure; molecular assembly /self assembly; molecular film; peptide chemical synthesis; phase change; phospholipids; protein folding; protein structure function; proteins; pulmonary surfactants; respiratory distress syndrome of newborn; serum albumin; surface property; synthetic protein; viscosity

Developing new surfactant substitutes requires (1) understanding the roles of the lipids and proteins in native lung surfactant; (2) developing easy to synthesize analogs of the native surfactant proteins; (3) optimizing lipid composition for low surface tension and rapid respreading and adsorption; (4) understanding surfactant inhibition by serum proteins and optimizing surfactant composition to minimize this inhibition. We will use modern biophysical techniques including Langmuir isotherms, fluorescence and Brewster angle optical microscopy, atomic force microscopy, viscometry, and x-ray diffraction to determine the phase behavior and morphology of lung surfactants. Specifically: (1) SP-B eliminates squeeze-out of unsaturated lipids and makes monolayer collapse more reversible. SP-C is more efficient at eliminating squeeze-out, and is less efficient at altering collapse. Is there an optimal ratio of SP-B to SP-C, or are the proteins interchangable? (2) Can we make a better SP-B and/or SP-C? Based on the known amino acid sequences of SP-B and SP-C, we will synthesize a family of homodimer peptides that mimic the characteristics of SP-B and SP-C. (3) Is there an optimal solid phase fraction in surfactant monolayers? Adding palmitic acid (PA) to DPPC influences the fluid to solid phase transitions and helps lower the surface tension. We plan to determine the composition of the solid phase in model surfactants and Survanta. (4) What leads to surfactant inhibition by serum proteins? Blood, plasma and serum proteins, lysolipids, and meconium may compete for interfacial area or might solubilize or degrade constituents of surfactant, thereby impairing monolayer function. We propose to investigate the interaction of model lung surfactant mixtures with human serum albumin to determine the molecular mechanisms of surfactant inhibition. (S) Can we relate monolayer viscosity to monolayer morphology and composition? We have built a magnetic needle viscometer to carry out systematic studies of LS monolayer shear viscoelasticity as a function of: i) protein content; ii) phase state of the monolayer. Concurrently, an existing double-barrier Langmuir trough will be adapted to do dilational viscoelasticity under the same conditions.

开发新的表面活性剂替代品需要（1）了解天然肺表面活性剂中脂质和蛋白质的作用;（2）开发易于合成的天然表面活性剂蛋白质类似物;（3）优化脂质组成，以实现低表面张力和快速再扩散和吸附;（4）了解血清蛋白对表面活性剂的抑制作用，并优化表面活性剂组成，以最大限度地减少这种抑制作用。我们将使用现代生物物理技术，包括朗缪尔等温线，荧光和布鲁斯特角光学显微镜，原子力显微镜，粘度计，和X-射线衍射，以确定肺表面活性剂的相行为和形态。具体而言：（1）SP-B消除了不饱和脂质的挤出，使单层塌陷更可逆。SP-C在消除挤出方面更有效，而在改变塌陷方面效率较低。SP-B与SP-C是否存在最佳比例，或者蛋白质是否可以互换？(2)我们能做一个更好的SP-B和/或SP-C吗？基于已知的SP-B和SP-C的氨基酸序列，我们将合成一个类似于SP-B和SP-C特征的同源二聚体肽家族。(3)表面活性剂单分子膜中是否存在最佳固相分数？向DPPC中添加棕榈酸（PA）影响流体到固体的相变并有助于降低表面张力。我们计划确定模型表面活性剂和Survanta中固相的组成。(4)是什么导致了血清蛋白对表面活性剂的抑制？血液、血浆和血清蛋白、溶血脂和胎粪可能会竞争界面面积，或者可能溶解或降解表面活性剂的成分，从而损害单层功能。我们建议研究模型肺表面活性物质混合物与人血清白蛋白的相互作用，以确定表面活性物质抑制的分子机制。(S)我们能否将单分子层粘度与单分子层形态和成分联系起来？我们建立了一个磁针粘度计进行系统的研究LS单分子膜剪切粘弹性的函数：i）蛋白质含量; ii）单分子膜的相状态。同时，现有的双势垒朗缪尔槽将在相同的条件下进行扩张粘弹性。