2NWD

Structure of chemically synthesized human lysozyme at 1 Angstrom resolution

2NWD の概要

• エントリーDOI: 10.2210/pdb2nwd/pdb
• 分子名称: Lysozyme C (2 entities in total)
• 機能のキーワード: native chemical ligation, chemical protein synthesis, convergent synthesis, hydrolase
• 細胞内の位置: Secreted: P61626
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 14720.69

構造登録者

Durek, T.,Torbeev, V.Y.,Kent, S.B.H. (登録日: 2006-11-14, 公開日: 2006-12-19, 最終更新日: 2024-10-30)

主引用文献

Durek, T.,Torbeev, V.Y.,Kent, S.B.
Convergent chemical synthesis and high-resolution x-ray structure of human lysozyme.
Proc.Natl.Acad.Sci.Usa, 104:4846-4851, 2007

PubMed Abstract: In this article, we report the total chemical synthesis of human lysozyme. Lysozyme serves as a widespread model system in various fields of biochemical research, including protein folding, enzyme catalysis, and amyloidogenesis. The 130-aa wild-type polypeptide chain of the human enzyme was assembled from four polypeptide segments by using native chemical ligation in a fully convergent fashion. Key to the assembly strategy is the application of the recently developed kinetically controlled ligation methodology, which provides efficient control over the ligation of two peptide (alpha)thioesters to yield a unique product. This result enables the facile preparation of a 64-residue peptide (alpha)thioester; this segment is joined by native chemical ligation to a 66-aa Cys peptide, to yield the target 130-aa polypeptide chain. The synthetic polypeptide chain was folded in vitro into a defined tertiary structure with concomitant formation of four disulfides, as shown by 2D TOCSY NMR spectroscopy. The structure of the synthetic human lysozyme was confirmed by high-resolution x-ray diffraction, giving the highest-resolution structure (1.04 A) observed to date for this enzyme. Synthetic lysozyme was obtained in good yield and excellent purity and had full enzymatic activity. This facile and efficient convergent synthesis scheme will enable preparation of unique chemical analogs of the lysozyme molecule and will prove useful in numerous areas of lysozyme research in the future.

PubMed: 17360367
DOI: 10.1073/pnas.0610630104

実験手法

X-RAY DIFFRACTION (1.04 Å)