Yu Peng, Liu Bo, Kodadek Thomas
Department of Internal Medicine, University of Texas Southwestern Medical Cetner, 5323 Harry Hines Blvd., Dallas, Texas 75390-8573.
Nat Biotechnol. 2005 Jun;23(6):746-51. doi: 10.1038/nbt1099. Epub 2005 May 22.
There is great interest in the identification of synthetic molecules that are capable of manipulating protein-protein interactions in living cells. Peptides, unlike other classes of small molecules, have binding properties appropriate for this application, but most are poorly cell permeable and sensitive to proteases. Therefore, considerable effort has been expended in the development of libraries of oligomeric peptide-like molecules. However, there are no clear-cut rules to guide the design of libraries rich in cell permeable compounds. Furthermore, currently available empirical methods to assess permeability may not accurately reflect true permeability and/or are capable of only modest throughput. We describe here an assay for assessing the relative cell permeability of synthetic molecules in the context of steroid fusions that is capable of high throughput and can be used in any transfectable cell line.
人们对能够在活细胞中操纵蛋白质-蛋白质相互作用的合成分子的鉴定有着浓厚的兴趣。与其他小分子类别不同，肽具有适合此应用的结合特性，但大多数肽的细胞渗透性差且对蛋白酶敏感。因此，在开发寡聚肽样分子文库方面已经投入了大量精力。然而，目前尚无明确的规则来指导富含细胞可渗透化合物的文库设计。此外，目前可用的评估渗透性的经验方法可能无法准确反映真实的渗透性和/或通量有限。我们在此描述一种在类固醇融合背景下评估合成分子相对细胞渗透性的测定方法，该方法具有高通量，可用于任何可转染的细胞系。
