Latge Cristiane, Cabral Katia M S, de Oliveira Guilherme A P, Raymundo Diana P, Freitas Julia A, Johanson Laizes, Romão Luciana F, Palhano Fernando L, Herrmann Torsten, Almeida Marcius S, Foguel Debora
From the Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21.941-902, Brazil.
the UFRJ/Pólo Xerém, Universidade Federal do Rio de Janeiro 25245-390, Rio de Janeiro, Brazil.
J Biol Chem. 2015 Aug 14;290(33):20527-40. doi: 10.1074/jbc.M115.662254. Epub 2015 Jul 6.
Cerebral dopamine neurotrophic factor (CDNF) is a promising therapeutic agent for Parkinson disease. As such, there has been great interest in studying its mode of action, which remains unknown. The three-dimensional crystal structure of the N terminus (residues 9-107) of CDNF has been determined, but there have been no published structural studies on the full-length protein due to proteolysis of its C-terminal domain, which is considered intrinsically disordered. An improved purification protocol enabled us to obtain active full-length CDNF and to determine its three-dimensional structure in solution. CDNF contains two well folded domains (residues 10-100 and 111-157) that are linked by a loop of intermediate flexibility. We identified two surface patches on the N-terminal domain that were characterized by increased conformational dynamics that should allow them to embrace active sites. One of these patches is formed by residues Ser-33, Leu-34, Ala-66, Lys-68, Ile-69, Leu-70, Ser-71, and Glu-72. The other includes a flexibly disordered N-terminal tail (residues 1-9), followed by the N-terminal portion of α-helix 1 (residues Cys-11, Glu-12, Val-13, Lys-15, and Glu-16) and residue Glu-88. The surface of the C-terminal domain contains two conserved active sites, which have previously been identified in mesencephalic astrocyte-derived neurotrophic factor, a CDNF paralog, which corresponds to its intracellular mode of action. We also showed that CDNF was able to protect dopaminergic neurons against injury caused by α-synuclein oligomers. This advises its use against physiological damages caused by α-synuclein oligomers, as observed in Parkinson disease and several other neurodegenerative diseases.
脑源性多巴胺神经营养因子（CDNF）是一种很有前景的帕金森病治疗药物。因此，人们对研究其作用方式产生了浓厚兴趣，但其作用方式仍不清楚。已确定了CDNF N端（第9 - 107位氨基酸残基）的三维晶体结构，但由于其C端结构域被认为是内在无序的且会发生蛋白水解，所以尚未有关于全长蛋白的结构研究报道。一种改进的纯化方案使我们能够获得有活性的全长CDNF，并确定其在溶液中的三维结构。CDNF包含两个折叠良好的结构域（第10 - 100位和第111 - 157位氨基酸残基），它们通过一个具有中等柔韧性的环相连。我们在N端结构域上鉴定出两个表面区域，其特征是构象动力学增加，这应使其能够容纳活性位点。其中一个区域由Ser - 33、Leu - 34、Ala - 66、Lys - 68、Ile - 69、Leu - 70、Ser - 71和Glu - 72这些氨基酸残基形成。另一个区域包括一个灵活无序的N端尾巴（第1 - 9位氨基酸残基），接着是α - 螺旋1的N端部分（Cys - 11、Glu - 12、Val - 13、Lys - 15和Glu - 16这些氨基酸残基）以及Glu - 88氨基酸残基。C端结构域的表面包含两个保守的活性位点，这些位点先前已在中脑星形胶质细胞源性神经营养因子（一种CDNF旁系同源物）中鉴定出来，这与其细胞内作用方式相对应。我们还表明，CDNF能够保护多巴胺能神经元免受α - 突触核蛋白寡聚体造成的损伤。这表明它可用于对抗帕金森病和其他几种神经退行性疾病中观察到的由α - 突触核蛋白寡聚体引起的生理损伤。
