Dietrich C, Schmitt L, Tampé R
Lehrstuhl für Biophysik E22, Technische Universität München, Garching, Germany.
Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9014-8. doi: 10.1073/pnas.92.20.9014.
In molecular biology, the expression of fusion proteins is a very useful and well-established technique for the identification and one-step purification of gene products. Even a short fused sequence of five or six histidines enables proteins to bind to an immobilized metal ion chelate complex. By synthesis of a class of chelator lipids, we have transferred this approach to the concept of self-assembly. The specific interaction and lateral organization of a fluorescent fusion molecule containing a C-terminal oligohistidine sequence was studied by film balance techniques in combination with epifluorescence microscopy. Due to the phase behavior of the various lipid mixtures used, the chelator lipids can be laterally structured, generating two-dimensional arrays of histidine-tagged biomolecules. Because of the large variety of fusion proteins already available, this concept represents a powerful technique for orientation and organization of proteins at lipid interfaces with applications in biosensing, biofunctionalization of nanostructured interfaces, two-dimensional crystallization, and studies of lipid-anchored proteins.
在分子生物学中，融合蛋白的表达是一种非常有用且成熟的技术，可用于基因产物的鉴定和一步纯化。即使是短短五六个组氨酸的融合序列，也能使蛋白质与固定化金属离子螯合物结合。通过合成一类螯合脂质，我们已将这种方法应用于自组装概念。采用膜天平技术结合落射荧光显微镜，研究了含有C端寡聚组氨酸序列的荧光融合分子的特异性相互作用和侧向组织。由于所使用的各种脂质混合物的相行为，螯合脂质可以形成侧向结构，产生组氨酸标记生物分子的二维阵列。鉴于已有大量融合蛋白，这一概念代表了一种强大的技术，可用于在脂质界面上对蛋白质进行定向和组织，应用于生物传感、纳米结构界面的生物功能化、二维结晶以及脂质锚定蛋白的研究。
