1K3Q

NMR structure of the FHA1 Domain of Rad53 in Complex with a Rad9-derived Phosphothreonine (at T192) Peptide

1K3Q の概要

• エントリーDOI: 10.2210/pdb1k3q/pdb
• 関連するPDBエントリー: 1G3G 1K3J 1K3N
• 分子名称: Protein Kinase SPK1, DNA repair protein Rad9 (2 entities in total)
• 機能のキーワード: fha domain, rad53, rad9, phosphothreonine, phosphoprotein, transferase-cell cycle complex, transferase/cell cycle
• 由来する生物種: Saccharomyces cerevisiae (baker's yeast); 詳細
• 細胞内の位置: Nucleus: P22216 P14737
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 18582.96

構造登録者

Yuan, C.,Yongkiettrakul, S.,Byeon, I.-J.L.,Zhou, S.,Tsai, M.-D. (登録日: 2001-10-03, 公開日: 2001-12-05, 最終更新日: 2024-10-30)

主引用文献

Yuan, C.,Yongkiettrakul, S.,Byeon, I.J.,Zhou, S.,Tsai, M.D.
Solution structures of two FHA1-phosphothreonine peptide complexes provide insight into the structural basis of the ligand specificity of FHA1 from yeast Rad53.
J.Mol.Biol., 314:563-575, 2001

PubMed Abstract: Rad53, a yeast checkpoint protein involved in regulating the repair of DNA damage, contains two forkhead-associated domains, FHA1 and FHA2. Previous combinatorial library screening has shown that FHA1 strongly selects peptides containing a pTXXD motif. Subsequent location of this motif within the sequence of Rad9, the target protein, coupled with spectroscopic analysis has led to identification of a tight binding sequence that is likely the binding site of FHA1: (188)SLEV(pT)EADATFVQ(200). We present solution structures of FHA1 in complex with this pT-peptide and with another Rad9-derived pT-peptide that has ca 30-fold lower affinity, (148)KKMTFQ(pT)PTDPLE(160). Both complexes showed intermolecular NOEs predominantly between three peptide residues (pT, +1, and +2 residues) and five FHA1 residues (S82, R83, S85, T106, and N107). Furthermore, the following interactions were implicated on the basis of chemical shift perturbations and structural analysis: the phosphate group of the pT residue with the side-chain amide group of N86 and the guanidino group of R70, and the carboxylate group of Asp (at the +3 position) with the guanidino group of R83. The generated structures revealed a similar binding mode adopted by these two peptides, suggesting that pT and the +3 residue Asp are the major contributors to binding affinity and specificity, while +1 and +2 residues could provide additional fine-tuning. It was also shown that FHA1 does not bind to the corresponding pS-peptides or a related pY-peptide. We suggest that differentiation between pT and pS-peptides by FHA1 can be attributed to hydrophobic interactions between the methyl group of the pT residue and the aliphatic protons of R83, S85, and T106 from FHA1.

PubMed: 11846567
DOI: 10.1006/jmbi.2001.5140

実験手法

SOLUTION NMR