Miyamoto Kazuhide, Uechi Airi, Saito Kazuki
Department of Pharmaceutical Health Care, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, Hyogo, Japan.
Protein Sci. 2017 Aug;26(8):1681-1686. doi: 10.1002/pro.3201. Epub 2017 Jun 11.
Human RING finger protein 141 (RFP141) is a germ cell-specific transcription factor during spermatogenesis. We synthesized a compact construct encoding the C-terminal zinc finger of RFP141 (RFP141C peptide). Herein we determined the solution structure of the RFP141C peptide by nuclear magnetic resonance (NMR). Moreover, NMR data and the chemical modification of cysteine residues demonstrated that the RFP141C peptide binds to two zinc atoms in a cross-brace arrangement. The Simple Modular Architecture Research Tool database predicted the structure of RFP141C as a RING finger. However, the actual structure of the RFP141C peptide adopts an atypical compact C HC -type RING fold. The position and range of the helical active site of the RFP141C structure were elucidated at the atomic level. Therefore, structural analysis may allow RFP141C to be used for designing an artificial RING finger possessing specific ubiquitin-conjugating enzyme (E2)-binding capabilities.
人类环状结构域蛋白141（RFP141）是精子发生过程中一种生殖细胞特异性转录因子。我们合成了一种编码RFP141 C末端锌指的紧凑构建体（RFP141C肽）。在此，我们通过核磁共振（NMR）确定了RFP141C肽的溶液结构。此外，NMR数据和半胱氨酸残基的化学修饰表明，RFP141C肽以交叉支撑排列与两个锌原子结合。简单模块化结构研究工具数据库将RFP141C的结构预测为一种环状结构域。然而，RFP141C肽的实际结构采用了非典型的紧凑C HC型环状折叠。RFP141C结构的螺旋活性位点的位置和范围在原子水平上得到了阐明。因此，结构分析可能使RFP141C可用于设计具有特定泛素结合酶（E2）结合能力的人工环状结构域。
