BeF3- acts as a phosphate analog in proteins phosphorylated on aspartate:: Structure of a BeF3- complex with phosphoserine phosphatase
BeF3- acts as a phosphate analog in proteins phosphorylated on aspartate:: Structure of a BeF3- complex with phosphoserine phosphatase
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DOI：
10.1073/pnas.131213698
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发表时间：
2001-07-17
影响因子：
11.1
通讯作者：
Yan, DL
中科院分区：
文献类型：
--
作者：
Cho, H;Wang, WR;Yan, DL
蛋白质磷酸天冬氨酸键发挥多种作用。在双组分信号转导系统的响应调节蛋白中，天冬氨酸残基的磷酸化与从非活性构象到活性构象的变化相结合。在卤酸脱卤酶 (HAD) 超家族的磷酸酶和变位酶中，磷酸天冬氨酸充当磷酸转移反应的中间体，而在同样属于 HAD 家族成员的 P 型 ATP 酶中，它负责将化学能转换为离子梯度。在每种情况下，磷酸天冬氨酸键的不稳定性都阻碍了对磷酸化形式的详细研究。对于反应调节剂来说，最近通过磷酸盐类似物 BeF3- 克服了这一困难，BeF3- 与受体域的活性位点天冬氨酸产生持久的复合物。现在，我们通过以 1.5 埃分辨率解析 BeF3- 与来自詹氏甲烷球菌的磷酸丝氨酸磷酸酶 (PSP) 复合物的 X 射线晶体结构，将该类似物的应用扩展到 HAD 超家族成员。该结构与磷酸酶中间体的结构相当：BeF3- 通过磷酰基的四面体几何结构与 Asp-II 结合，与 Mg2+ 配位，并与 HAD 超家族中保守的活性位点周围的残基结合。 BeF3-活性位点的比较。 PSP 和 BeF3-。 CeY 是一种受体域/反应调节因子，它揭示了惊人的相似性，不仅可以深入了解 PSP 的功能，还可以深入了解 P 型 ATP 酶的功能。我们的结果表明，使用 BeF3- 对形成磷酸天冬氨酸键的蛋白质进行结构研究将远远超出反应调节剂的范围。
Protein phosphoaspartate bonds play a variety of roles. In response regulator proteins of two-component signal transduction systems, phosphorylation of an aspartate residue is coupled to a change from an inactive to an active conformation. In phosphatases and mutases of the haloacid dehalogenase (HAD) superfamily, phosphoaspartate serves as an intermediate in phosphotransfer reactions, and in P-type ATPases, also members of the HAD family, it serves in the conversion of chemical energy to ion gradients. In each case, lability of the phosphoaspartate linkage has hampered a detailed study of the phosphorylated form. For response regulators, this difficulty was recently overcome with a phosphate analog, BeF3-, which yields persistent complexes with the active site aspartate of their receiver domains. We now extend the application of this analog to a HAD superfamily member by solving at 1.5-Angstrom resolution the x-ray crystal structure of the complex of BeF3- with phosphoserine phosphatase (PSP) from Methanococcus jannaschii. The structure is comparable to that of a phosphoenzyme intermediate: BeF3- is bound to Asp-ll with the tetrahedral geometry of a phosphoryl group, is coordinated to Mg2+, and is bound to residues surrounding the active site that are conserved in the HAD superfamily. Comparison of the active sites of BeF3-. PSP and BeF3-. CeY, a receiver domain/response regulator, reveals striking similarities that provide insights into the function not only of PSP but also of P-type ATPases. Our results indicate that use of BeF3- for structural studies of proteins that form phosphoaspartate linkages will extend well beyond response regulators.