Krakowiak Agnieszka, Kocoń-Rębowska Beata, Dolot Rafał, Piotrzkowska Danuta
Department of Bioorganic Chemistry, Polish Academy of Sciences, Centre of Molecular and Macromolecular Studies, Lodz, Poland.
FEBS Lett. 2017 Feb;591(3):548-559. doi: 10.1002/1873-3468.12560. Epub 2017 Feb 5.
Fragile histidine triad protein (Fhit) is a protein which primarily hydrolyses dinucleoside polyphosphates. To investigate possible interactions between the protein and a substrate, we used a nonhydrolyzable phosphorothioate analog of Ap A, containing 5-bromo-2'-deoxyuridine instead of one adenosine residue. Photocrosslinking, followed by LC-MS experiments, determined a complex in which the probe was covalently linked to the NDSIYEELQK peptide (residues 110-119). The peptide was located within the 'disordered' region, which is invisible in the known crystal structures of Fhit. This invisible and flexible part seems to play a role in the stabilization of the Fhit-substrate complex, which may be important for its tumor suppressor activity.
脆性组氨酸三联体蛋白（Fhit）是一种主要水解二核苷多磷酸的蛋白质。为了研究该蛋白质与底物之间可能存在的相互作用，我们使用了ApA的一种不可水解的硫代磷酸酯类似物，其中一个腺苷残基被5-溴-2'-脱氧尿苷取代。通过光交联，随后进行液相色谱-质谱实验，确定了一种复合物，其中探针与NDSIYEELQK肽（第110-119位残基）共价连接。该肽位于“无序”区域内，在Fhit已知的晶体结构中不可见。这个不可见且灵活的部分似乎在Fhit-底物复合物的稳定中发挥作用，这可能对其肿瘤抑制活性很重要。
