Department of Physiology and Pharmacology, University of Western Ontario, London, Ontario, Canada; Department of Burns and Cutaneous Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, China.
J Cell Physiol. 2014 Sep;229(9):1121-9. doi: 10.1002/jcp.24630.
There is a critical need for techniques that directly monitor protein synthesis within cells isolated from normal and diseased tissue. Fibrotic disease, for which there is no drug treatment, is characterized by the overexpression of collagens. Here, we use a bioinformatics approach to identify a pair of glycine and proline isoacceptor tRNAs as being specific for the decoding of collagen mRNAs, leading to development of a FRET-based approach, dicodon monitoring of protein synthesis (DiCoMPS), that directly monitors the synthesis of collagen. DiCoMPS aimed at detecting collagen synthesis will be helpful in identifying novel anti-fibrotic compounds in cells derived from patients with fibrosis of any etiology, and, suitably adapted, should be widely applicable in monitoring the synthesis of other proteins in cells.
目前非常需要能够直接监测从正常和病变组织中分离出的细胞内蛋白质合成的技术。纤维化疾病是一种没有药物治疗的疾病，其特征是胶原蛋白过度表达。在这里，我们使用生物信息学方法来鉴定一对甘氨酸和脯氨酸同功受体 tRNA，它们是专门为解码胶原蛋白 mRNA 而设计的，从而开发了一种基于 FRET 的方法，即二密码子监测蛋白质合成（DiCoMPS），该方法可以直接监测胶原蛋白的合成。DiCoMPS 旨在检测胶原蛋白的合成，这将有助于在任何病因引起的纤维化患者衍生的细胞中识别新型抗纤维化化合物，并且在适当调整后，应该可以广泛应用于监测细胞中其他蛋白质的合成。
