1OID

5'-Nucleotidase (E. coli) with an Engineered Disulfide Bridge (S228C, P513C)

1OID の概要

• エントリーDOI: 10.2210/pdb1oid/pdb
• 関連するPDBエントリー: 1HO5 1HP1 1HPU 1OI8 1OIE 1USH 2USH
• 分子名称: PROTEIN USHA, NICKEL (II) ION (3 entities in total)
• 機能のキーワード: metalloprotein, hydrolase, domain movement, disulfide engineering, udp-sugar hydrolase, conformational trapping
• 由来する生物種: ESCHERICHIA COLI
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 118574.10

構造登録者

Schultz-Heienbrok, R.,Maier, T.,Straeter, N. (登録日: 2003-06-13, 公開日: 2004-06-10, 最終更新日: 2024-11-06)

主引用文献

Schultz-Heienbrok, R.,Maier, T.,Straeter, N.
Trapping a 96 Degree Domain Rotation in Two Distinct Conformations by Engineered Disulfide Bridges
Protein Sci., 13:1811-, 2004

PubMed Abstract: Engineering disulfide bridges is a common technique to lock a protein movement in a defined conformational state. We have designed two double mutants of Escherichia coli 5'-nucleotidase to trap the enzyme in both an open (S228C, P513C) and a closed (P90C, L424C) conformation by the formation of disulfide bridges. The mutant proteins have been expressed, purified, and crystallized, to structurally characterize the designed variants. The S228C, P513C is a double mutant crystallized in two different crystal forms with three independent conformers, which differ from each other by a rotation of up to 12 degrees of the C-terminal domain with respect to the N-terminal domain. This finding, as well as an analysis of the domain motion in the crystal, indicates that the enzyme still exhibits considerable residual domain flexibility. In the double mutant that was designed to trap the enzyme in the closed conformation, the structure analysis reveals an unexpected intermediate conformation along the 96 degrees rotation trajectory between the open and closed enzyme forms. A comparison of the five independent conformers analyzed in this study shows that the domain movement of the variant enzymes is characterized by a sliding movement of the residues of the domain interface along the interface, which is in contrast to a classical closure motion where the residues of the domain interface move perpendicular to the interface.

PubMed: 15215524
DOI: 10.1110/PS.04629604

実験手法

X-RAY DIFFRACTION (2.1 Å)