Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
Beryllium Discovery Corp., with present address of UCB Biosciences, Bedford, MA 01730, USA.
Structure. 2021 Aug 5;29(8):873-885.e5. doi: 10.1016/j.str.2021.03.008. Epub 2021 Mar 29.
Taspase1 is an Ntn-hydrolase overexpressed in primary human cancers, coordinating cancer cell proliferation, invasion, and metastasis. Loss of Taspase1 activity disrupts proliferation of human cancer cells in vitro and in mouse models of glioblastoma. Taspase1 is synthesized as an inactive proenzyme, becoming active upon intramolecular cleavage. The activation process changes the conformation of a long fragment at the C-terminus of the α subunit, for which no full-length structural information exists and whose function is poorly understood. We present a cloning strategy to generate a circularly permuted form of Taspase1 to determine the crystallographic structure of active Taspase1. We discovered that this region forms a long helix and is indispensable for the catalytic activity of Taspase1. Our study highlights the importance of this element for the enzymatic activity of Ntn-hydrolases, suggesting that it could be a potential target for the design of inhibitors with potential to be developed into anticancer therapeutics.
Taspase1 是一种在原发性人类癌症中过度表达的 Ntn-水解酶，它协调癌细胞的增殖、侵袭和转移。Taspase1 活性的丧失会破坏人类癌细胞在体外和胶质母细胞瘤小鼠模型中的增殖。Taspase1 作为无活性的前酶合成，在分子内切割后变得活跃。激活过程改变了 α 亚基 C 末端长片段的构象，目前尚无该片段全长结构信息，其功能也知之甚少。我们提出了一种克隆策略来产生 Taspase1 的环状排列形式，以确定活性 Taspase1 的晶体结构。我们发现该区域形成一个长螺旋，并对 Taspase1 的催化活性是不可或缺的。我们的研究强调了该元件对 Ntn-水解酶酶活性的重要性，表明它可能是设计抑制剂的潜在靶点，具有开发成抗癌治疗药物的潜力。
