PDF(Pubmed)
Abstract：
Solid tumours can universally evade contact inhibition of proliferation (CIP), a mechanism halting cell proliferation when cell-cell contact occurs. Merlin, an ERM-like protein, crucially regulates CIP and is frequently deactivated in various cancers, indicating its significance as a tumour suppressor in cancer biology. Despite extensive investigations into Merlin\'s role in cancer, its lack of intrinsic catalytic activity and frequent conformation changes have made it notoriously challenging to study. To address this challenge, we harnessed innovative luciferase technologies to create and validate a NanoBiT split-luciferase biosensor system in which Merlin is cloned between two split components (LgBiT and SmBiT) of NanoLuc luciferase. This system enables precise quantification of Merlin\'s conformation and activity both in vitro and within living cells. This biosensor significantly enhances the study of Merlin\'s molecular functions, serving as a potent tool for exploring its contributions to CIP and tumorigenesis.
摘要：
实体肿瘤可以普遍逃避增殖接触抑制（CIP），一种机制，当细胞与细胞接触时停止细胞增殖。梅林,ERM样蛋白质,至关重要地调节CIP，并且在各种癌症中经常失活，表明其在癌症生物学中作为肿瘤抑制因子的意义。尽管对Merlin在癌症中的作用进行了广泛的调查，它缺乏内在的催化活性和频繁的构象变化使其研究具有挑战性。为了应对这一挑战，我们利用创新的荧光素酶技术来创建和验证NanoBiT分裂荧光素酶生物传感器系统，其中Merlin被克隆在NanoLuc荧光素酶的两个分裂成分（LgBiT和SmBiT）之间。该系统能够在体外和活细胞内精确定量Merlin的构象和活性。这种生物传感器显着增强了对Merlin分子功能的研究，作为探索其对CIP和肿瘤发生的贡献的有力工具。
