Ciancaglini P, Simão A M S, Bolean M, Millán J L, Rigos C F, Yoneda J S, Colhone M C, Stabeli R G
Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto da (FFCLRP), Universidade de São Paulo - USP, 14040-901, Ribeirão Preto, SP, Brazil.
Sanford Children's Health Research Center, Sanford - Burnham Medical Research Institute, La Jolla, CA, USA.
Biophys Rev. 2012 Mar;4(1):67-81. doi: 10.1007/s12551-011-0065-4. Epub 2012 Jan 18.
Proteoliposomes are systems that mimic lipid membranes (liposomes) to which a protein has been incorporated or inserted. During the last decade, these systems have gained prominence as tools for biophysical studies on lipid-protein interactions as well as for their biotechnological applications. Proteoliposomes have a major advantage when compared with natural membrane systems, since they can be obtained with a smaller number of lipidic (and protein) components, facilitating the design and interpretation of certain experiments. However, they have the disadvantage of requiring methodological standardization for incorporation of each specific protein, and the need to verify that the reconstitution procedure has yielded the correct orientation of the protein in the proteoliposome system with recovery of its functional activity. In this review, we chose two proteins under study in our laboratory to exemplify the steps necessary for the standardization of the reconstitution of membrane proteins in liposome systems: (1) alkaline phosphatase, a protein with a glycosylphosphatidylinositol anchor, and (2) Na,K-ATPase, an integral membrane protein. In these examples, we focus on the production of the specific proteoliposomes, as well as on their biochemical and biophysical characterization, with emphasis on studies of lipid-protein interactions. We conclude the chapter by highlighting current prospects of this technology for biotechnological applications, including the construction of nanosensors and of a multi-protein nanovesicular biomimetic to study the processes of initiation of skeletal mineralization.
蛋白脂质体是一种模拟脂质膜（脂质体）的体系，其中已掺入或插入了蛋白质。在过去十年中，这些体系作为研究脂质 - 蛋白质相互作用的生物物理工具及其生物技术应用而备受关注。与天然膜系统相比，蛋白脂质体具有一个主要优势，因为它们可以通过较少数量的脂质（和蛋白质）成分获得，便于某些实验的设计和解释。然而，它们的缺点是每种特定蛋白质的掺入都需要方法标准化，并且需要验证重组过程是否在蛋白脂质体系统中使蛋白质具有正确的方向并恢复其功能活性。在本综述中，我们选择了我们实验室正在研究的两种蛋白质，以举例说明脂质体系统中膜蛋白重组标准化所需的步骤：（1）碱性磷酸酶，一种具有糖基磷脂酰肌醇锚定的蛋白质，以及（2）钠钾 - ATP酶，一种整合膜蛋白。在这些例子中，我们重点关注特定蛋白脂质体的制备及其生化和生物物理特性，重点是脂质 - 蛋白质相互作用的研究。我们在本章结尾强调了该技术在生物技术应用方面的当前前景，包括构建纳米传感器和一种多蛋白纳米囊泡仿生体以研究骨骼矿化起始过程。
