Rabuka David, Forstner Martin B, Groves Jay T, Bertozzi Carolyn R
Department of Chemistry, University of California, Berkeley, California 94720, USA.
J Am Chem Soc. 2008 May 7;130(18):5947-53. doi: 10.1021/ja710644g. Epub 2008 Apr 11.
The controlled addition of structurally defined components to live cell membranes can facilitate the molecular level analysis of cell surface phenomena. Here we demonstrate that cell surfaces can be engineered to display synthetic bioactive polymers at defined densities by exogenous membrane insertion. The polymers were designed to mimic native cell-surface mucin glycoproteins, which are defined by their dense glycosylation patterns and rod-like structures. End-functionalization with a hydrophobic anchor permitted incorporation into the membranes of live cultured cells. We probed the dynamic behavior of cell-bound glycopolymers bearing various hydrophobic anchors and glycan structures using fluorescence correlation spectroscopy (FCS). Their diffusion properties mirrored those of many natural membrane-associated biomolecules. Furthermore, the membrane-bound glycopolymers were internalized into early endosomes similarly to endogenous membrane components and were capable of specific interactions with protein receptors. This system provides a platform to study cell-surface phenomena with a degree of chemical control that cannot be achieved using conventional biological tools.
向活细胞膜中可控地添加结构明确的成分有助于在分子水平上分析细胞表面现象。在此,我们证明通过外源膜插入可对细胞表面进行工程改造,以特定密度展示合成生物活性聚合物。这些聚合物被设计成模仿天然细胞表面粘蛋白糖蛋白,其由密集的糖基化模式和棒状结构所定义。用疏水锚进行末端功能化可使其掺入活培养细胞的膜中。我们使用荧光相关光谱法(FCS)探测了带有各种疏水锚和聚糖结构的细胞结合糖聚合物的动态行为。它们的扩散特性反映了许多天然膜相关生物分子的扩散特性。此外,膜结合糖聚合物与内源性膜成分类似地被内化到早期内体中,并且能够与蛋白质受体发生特异性相互作用。该系统提供了一个平台,可在一定程度的化学控制下研究细胞表面现象,这是使用传统生物学工具无法实现的。