Biotechnology Center, University of Technology Dresden, Dresden, Germany.
Proteomics. 2010 Apr;10(7):1455-62. doi: 10.1002/pmic.200900724.
To control their attachment to substrates and other cells, cells regulate their adhesion receptors. One regulatory process is receptor crosstalk, where the binding of one type of cell adhesion molecule influences the activity of another type. To identify such crosstalk and gain insight into their mechanisms, we developed the stimulated single-cell force spectroscopy assay. In this assay, the influence of a cells adhesion to one substrate on the strength of its adhesion to a second substrate is examined. The assay quantifies the adhesion of the cell and the contributions of specific adhesion receptors. This allows mechanisms by which the adhesion is regulated to be determined. Using the assay we identified crosstalk between collagen-binding integrin alpha(1)beta(1) and fibronectin-binding integrin alpha(5)beta(1) in HeLa cells. This crosstalk was unidirectional, from integrin alpha(1)beta(1) to integrin alpha(5)beta(1), and functioned by regulating the endocytosis of integrin alpha(5)beta(1). The single-cell assay should be expandable for the screening and quantification of crosstalk between various cell adhesion molecules and other cell surface receptors.
为了控制细胞对底物和其他细胞的附着，细胞会调节其粘附受体。一种调节过程是受体串扰，其中一种类型的细胞粘附分子的结合会影响另一种类型的活性。为了识别这种串扰并深入了解其机制，我们开发了受刺激的单细胞力谱测定法。在该测定中，检查了细胞对一种底物的粘附对其对第二种底物的粘附强度的影响。该测定法定量了细胞的粘附以及特定粘附受体的贡献。这可以确定调节粘附的机制。使用该测定法，我们在 HeLa 细胞中鉴定了胶原结合整合素α 1β 1 和纤连蛋白结合整合素α 5β 1 之间的串扰。这种串扰是单向的，从整合素α 1β 1 到整合素α 5β 1，通过调节整合素α 5β 1 的内吞作用起作用。单细胞测定法应该可以扩展用于筛选和定量各种细胞粘附分子和其他细胞表面受体之间的串扰。
