2CW1

Solution structure of the de novo-designed lambda Cro fold protein

2CW1 の概要

• エントリーDOI: 10.2210/pdb2cw1/pdb
• 分子名称: SN4m (1 entity in total)
• 機能のキーワード: lambda cro fold, de novo protein
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 7488.70

構造登録者

Isogai, Y.,Ito, Y.,Ikeya, T.,Shiro, Y.,Ota, M. (登録日: 2005-06-15, 公開日: 2005-12-13, 最終更新日: 2024-05-29)

主引用文献

Isogai, Y.,Ito, Y.,Ikeya, T.,Shiro, Y.,Ota, M.
Design of lambda Cro fold: solution structure of a monomeric variant of the de novo protein.
J.Mol.Biol., 354:801-814, 2005

PubMed Abstract: One of the classical DNA-binding proteins, bacteriophage lambda Cro, forms a homodimer with a unique fold of alpha-helices and beta-sheets. We have computationally designed an artificial sequence of 60 amino acid residues to stabilize the backbone tertiary structure of the lambda Cro dimer by simulated annealing using knowledge-based structure-sequence compatibility functions. The designed amino acid sequence has 25% identity with that of natural lambda Cro and preserves Phe58, which is important for formation of the stably folded structure of lambda Cro. The designed dimer protein and its monomeric variant, which was redesigned by the insertion of a beta-hairpin sequence at the C-terminal region to prevent dimerization, were synthesized and biochemically characterized to be well folded. The designed protein was monomeric under a wide range of protein concentrations and its solution structure was determined by NMR spectroscopy. The solved structure is similar to that of a monomeric variant of natural lambda Cro with a root-mean-square deviation of the polypeptide backbones at 2.1A and has a well-packed protein core. Thus, our knowledge-based functions provide approximate but essential relationships between amino acid sequences and protein structures, and are useful for finding novel sequences that are foldable into a given target structure.

PubMed: 16289118
DOI: 10.1016/j.jmb.2005.10.005

実験手法

SOLUTION NMR