Biologics Discovery, Bristol-Myers Squibb Research and Development, 100 Binney Street, Cambridge, MA 02142, USA.
Molecular Structure and Design, Bristol-Myers Squibb Research and Development, 5 Research Parkway, Wallingford, CT 06492, USA.
Acta Crystallogr F Struct Biol Commun. 2021 Jan 1;77(Pt 1):22-28. doi: 10.1107/S2053230X20016015. Epub 2020 Dec 22.
Hematopoietic progenitor kinase 1 (HPK1) is an intracellular kinase that plays an important role in modulating tumor immune response and thus is an attractive target for drug discovery. Crystallization of the wild-type HPK1 kinase domain has been hampered by poor expression in recombinant systems and poor solubility. In this study, yeast surface display was applied to a library of HPK1 kinase-domain variants in order to select variants with an improved expression level and solubility. The HPK1 variant with the most improved properties contained two mutations, crystallized readily in complex with several small-molecule inhibitors and provided valuable insight to guide structure-based drug design. This work exemplifies the benefit of yeast surface display towards engineering crystallizable proteins and thus enabling structure-based drug discovery.
造血祖细胞激酶 1（HPK1）是一种细胞内激酶，在调节肿瘤免疫反应方面发挥着重要作用，因此是药物发现的一个有吸引力的靶点。野生型 HPK1 激酶结构域的结晶受到重组系统中表达不佳和溶解度差的阻碍。在这项研究中，酵母表面展示被应用于 HPK1 激酶结构域变体文库，以选择具有改善表达水平和溶解度的变体。具有最佳性质的 HPK1 变体包含两个突变，与几种小分子抑制剂容易结晶，并为基于结构的药物设计提供了有价值的见解。这项工作例证了酵母表面展示在工程可结晶蛋白方面的优势，从而实现了基于结构的药物发现。
