1QVY

Crystal structure of RhoGDI K(199,200)R double mutant

1QVY の概要

• エントリーDOI: 10.2210/pdb1qvy/pdb
• 関連するPDBエントリー: 1KMT 1RHO
• 分子名称: Rho GDP-dissociation inhibitor 1, SULFATE ION (3 entities in total)
• 機能のキーワード: protein crystallization; rational surface mutagenesis; rhogdi; high resolution; x-ray diffraction, signaling protein inhibitor
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Cytoplasm: P52565
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 64869.58

構造登録者

Czepas, J.,Devedjiev, Y.,Krowarsh, D.,Derewenda, U.,Derewenda, Z.S. (登録日: 2003-08-29, 公開日: 2004-02-10, 最終更新日: 2023-08-16)

主引用文献

Czepas, J.,Devedjiev, Y.,Krowarsch, D.,Derewenda, U.,Otlewski, J.,Derewenda, Z.S.
The impact of Lys-->Arg surface mutations on the crystallization of the globular domain of RhoGDI.
Acta Crystallogr.,Sect.D, 60:275-280, 2004

PubMed Abstract: The potential of rational surface mutagenesis for enhanced protein crystallization is being probed in an ongoing effort. In previous work, it was hypothesized that residues with high conformational entropy such as Glu and Lys are suitable targets for surface mutagenesis, as they are rarely incorporated in crystal contacts or protein-protein interfaces. Previous experiments using Lys-->Ala, Glu-->Ala and Glu-->Asp mutants confirmed that mutated proteins were more likely to crystallize. In the present paper, the usefulness of Lys-->Arg mutations is studied. Several mutations of the globular domain of human RhoGDI were generated, including the single mutants K105R, K113R, K127R, K138R and K141R, the double mutants K(98,99)R and K(199,200)R and the triple mutants K(98,99,105)R and K(135,138,141)R. It is shown that Lys-->Arg mutants are more likely to crystallize than the wild-type protein, although not as likely as Lys-->Ala mutants. Out of the nine mutants tested, five produced diffracting crystals, including the K(199,200)R double mutant, which crystallized in a new space group and exceeded by approximately 1.0 A the resolution of the diffraction of the wild-type crystal. Major crystal contacts in the new lattice were created by the mutated epitope.

PubMed: 14747703
DOI: 10.1107/S0907444903026271

実験手法

X-RAY DIFFRACTION (1.6 Å)