Diemel R V, Bader D, Walch M, Hotter B, van Golde L M, Amann A, Haagsman H P, Putz G
Department of Anaesthesiology and Critical Care Medicine, The Leopold-Franzens-University of Innsbruck, Austria.
Arch Biochem Biophys. 2001 Jan 15;385(2):338-47. doi: 10.1006/abbi.2000.2157.
Surfactant protein B (SP-B) enhances lipid insertion into the alveolar air/liquid interface upon inhalation. The aim of this study was (i) to apply a palette of tests for a detailed biochemical and biophysical characterization of SP-B and (ii) to use these tests to compare native SP-B with a fluorescent (Bodipy) SP-B analog. The method of labeling was fast and resulted in a covalent fluorophore-protein bond. The ability of both proteins to spread a surfactant film on top of a buffer surface was determined in a spreading tray using the Wilhelmy plate technique to allow detection of alterations in surface tension and calculation of spreading velocities. In a captive bubble surfactometer surface tensions of spread films were measured. Similar biophysical properties were found for both native and Bodipy-labeled SP-B. It is concluded that the combination of tests used allows detection of small differences in structure and activity between the two proteins.
表面活性蛋白B（SP-B）在吸入时可增强脂质插入肺泡气/液界面的能力。本研究的目的是：（i）应用一系列测试对SP-B进行详细的生化和生物物理特性表征；（ii）使用这些测试比较天然SP-B与荧光（Bodipy）SP-B类似物。标记方法快速，且形成了共价荧光团-蛋白质键。使用Wilhelmy平板技术在铺展盘中测定了两种蛋白质在缓冲液表面上铺展表面活性剂膜的能力，以检测表面张力的变化并计算铺展速度。在俘获气泡表面张力仪中测量铺展膜的表面张力。天然SP-B和Bodipy标记的SP-B具有相似的生物物理特性。得出的结论是，所使用的测试组合能够检测出这两种蛋白质在结构和活性上的细微差异。
