The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA.
Quantum Tessera Consulting, LLC, Collegeville, PA 19426, USA.
Molecules. 2020 Apr 10;25(7):1760. doi: 10.3390/molecules25071760.
The human gamma-herpesviruses Epstein-Barr virus (EBV) (HHV-4) and Kaposi's sarcoma-associated herpesvirus (KSHV) (HHV-8) are responsible for a number of diseases, including various types of cancer. Epstein-Barr nuclear antigen 1 (EBNA1) from EBV and latency-associated nuclear antigen (LANA) from KSHV are viral-encoded DNA-binding proteins that are essential for the replication and maintenance of their respective viral genomes during latent, oncogenic infection. As such, EBNA1 and LANA are attractive targets for the development of small-molecule inhibitors. To this end, we performed a biophysical screen of EBNA1 and LANA using a fragment library by saturation transfer difference (STD)-NMR spectroscopy and surface plasmon resonance (SPR). We identified and validated a number of unique fragment hits that bind to EBNA1 or LANA. We also determined the high-resolution crystal structure of one fragment bound to EBNA1. Results from this screening cascade provide new chemical starting points for the further development of potent inhibitors for this class of viral proteins.
人类γ疱疹病毒 EBV（HHV-4）和卡波济肉瘤相关疱疹病毒 KSHV（HHV-8）可引起多种疾病，包括各种癌症。EBV 的 EBNA1 和 KSHV 的潜伏相关核抗原（LANA）是病毒编码的 DNA 结合蛋白，对于在潜伏、致癌感染期间复制和维持其各自的病毒基因组至关重要。因此，EBNA1 和 LANA 是开发小分子抑制剂的有吸引力的靶标。为此，我们使用片段文库通过饱和转移差异（STD）-NMR 光谱法和表面等离子体共振（SPR）对 EBNA1 和 LANA 进行了生物物理筛选。我们鉴定并验证了许多与 EBNA1 或 LANA 结合的独特片段。我们还确定了一个与 EBNA1 结合的片段的高分辨率晶体结构。该筛选级联的结果为进一步开发此类病毒蛋白的有效抑制剂提供了新的化学起点。
