4KVT

Crystal structure of a 6-helix coiled coil CC-Hex-L24C

4KVT の概要

• エントリーDOI: 10.2210/pdb4kvt/pdb
• 関連するPDBエントリー: 3R3K 4KVU 4KVV
• 分子名称: 6-helix coiled coil CC-Hex-L24C peptide (2 entities in total)
• 機能のキーワード: de novo coiled-coil assembly, de novo protein
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 20129.87

構造登録者

Burton, A.J.,Agnew, C.,Brady, R.L.,Woolfson, D.N. (登録日: 2013-05-23, 公開日: 2013-08-21, 最終更新日: 2024-10-30)

主引用文献

Burton, A.J.,Thomas, F.,Agnew, C.,Hudson, K.L.,Halford, S.E.,Brady, R.L.,Woolfson, D.N.
Accessibility, Reactivity, and Selectivity of Side Chains within a Channel of de Novo Peptide Assembly.
J.Am.Chem.Soc., 135:12524-12527, 2013

PubMed Abstract: Ab initio design of enzymes requires precise and predictable positioning of reactive functional groups within accessible and controlled environments of de novo protein scaffolds. Here we show that multiple thiol moieties can be placed within a central channel, with approximate dimensions 6 × 42 Å, of a de novo, six-helix peptide assembly (CC-Hex). Layers of six cysteine residues are introduced at two different sites ~6 (the "L24C" mutant) and ~17 Å (L17C) from the C-terminal opening of the channel. X-ray crystal structures confirm the mutant structures as hexamers with internal free thiol, rather than disulfide-linked cysteine residues. Both mutants are hexa-alkylated upon addition of iodoacetamide, demonstrating accessibility and full reactivity of the thiol groups. Comparison of the alkylation and unfolding rates of the hexamers indicates that access is directly through the channel and not via dissociation and unfolding of the assembly. Moreover, neither mutant reacts with iodoacetic acid, demonstrating selectivity of the largely hydrophobic channel. These studies show that it is possible to engineer reactive side chains with both precision and control into a de novo scaffold to produce protein-like structures with chemoselective reactivity.

PubMed: 23924058
DOI: 10.1021/ja4053027

実験手法

X-RAY DIFFRACTION (1.6 Å)