NMR study of the structure and dynamics of the BRCT domain from the kinetochore protein KKT4.
NMR study of the structure and dynamics of the BRCT domain from the kinetochore protein KKT4.
复制标题
对动粒蛋白 KKT4 的 BRCT 结构域的结构和动力学进行 NMR 研究。
DOI：
10.1007/s12104-024-10163-9
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发表时间：
2024
影响因子：
0.9
通讯作者：
Ludzia P
中科院分区：
文献类型：
--
作者：
Ludzia P
KKT4是一种多结构域的动粒蛋白，专为布氏锥虫等动体所特有。它与其他真核生物中已知的动粒蛋白缺乏显著的序列相似性。我们最近对KKT4的C-末端区域的X射线结构表明，它有一个串联的BRCT(BRCA1 C和Terminus)结构域折叠，在一个典型的磷酸化丝氨酸或苏氨酸的结合部位结合了一个硫酸盐离子。在这里，我们给出了KKT4(KKT4463-645)的BRCT结构域的1H，13C和15N共振指定。布鲁塞。我们证明了BRCT结构域可以利用X-射线结构中与硫酸盐离子结合的残基与溶液中的磷酸根离子结合。我们用这些赋值来表征溶液中BRCT结构域的二级结构和主干动力学。突变磷酸根离子结合INT所涉及的残基。BruceiKKT4 BRCT导致生长缺陷，证实了BRCT磷酸肽结合活性在体内的重要性。这些结果可能有助于未来合理的药物设计工作，以对抗动体寄生虫引起的疾病。
KKT4 is a multi-domain kinetochore protein specific to kinetoplastids, such asTrypanosoma brucei. It lacks significant sequence similarity to known kinetochore proteins in other eukaryotes. Our recent X-ray structure of the C-terminal region of KKT4 shows that it has a tandem BRCT (BRCA1 C Terminus) domain fold with a sulfate ion bound in a typical binding site for a phosphorylated serine or threonine. Here we present the1H,13C and15N resonance assignments for the BRCT domain of KKT4 (KKT4463–645) fromT. brucei. We show that the BRCT domain can bind phosphate ions in solution using residues involved in sulfate ion binding in the X-ray structure. We have used these assignments to characterise the secondary structure and backbone dynamics of the BRCT domain in solution. Mutating the residues involved in phosphate ion binding inT. bruceiKKT4 BRCT results in growth defects confirming the importance of the BRCT phosphopeptide-binding activity in vivo. These results may facilitate rational drug design efforts in the future to combat diseases caused by kinetoplastid parasites.