2JHT

CRYSTAL STRUCTURE OF RHOGDI K135T,K138T,K141T MUTANT

2JHT の概要

• エントリーDOI: 10.2210/pdb2jht/pdb
• 関連するPDBエントリー: 1CC0 1FSO 1FST 1FT0 1FT3 1HH4 1KMT 1QVY 1RHO 2BXW 2JHS 2JHU 2JHV 2JHW 2JHX 2JHY 2JHZ 2JI0
• 分子名称: RHO GDP-DISSOCIATION INHIBITOR 1, SULFATE ION, LITHIUM ION, ... (4 entities in total)
• 機能のキーワード: surface entropy reduction, inhibitor, gtpase activation, crystal engineering
• 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: 63046.35

構造登録者

Cooper, D.R.,Grelewska, K.,Derewenda, Z.S. (登録日: 2007-02-23, 公開日: 2007-05-08, 最終更新日: 2023-12-13)

主引用文献

Cooper, D.R.,Boczek, T.,Grelewska, K.,Pinkowska, M.,Sikorska, M.,Zawadzki, M.,Derewenda, Z.S.
Protein Crystallization by Surface Entropy Reduction: Optimization of the Ser Strategy
Acta Crystallogr.,Sect.D, 63:636-, 2007

PubMed Abstract: A strategy of rationally engineering protein surfaces with the aim of obtaining mutants that are distinctly more susceptible to crystallization than the wild-type protein has previously been suggested. The strategy relies on replacing small clusters of two to three surface residues characterized by high conformational entropy with alanines. This surface entropy reduction (or SER) method has proven to be an effective salvage pathway for proteins that are difficult to crystallize. Here, a systematic comparison of the efficacy of using Ala, His, Ser, Thr and Tyr to replace high-entropy residues is reported. A total of 40 mutants were generated and screened using two different procedures. The results reaffirm that alanine is a particularly good choice for a replacement residue and identify tyrosines and threonines as additional candidates that have considerable potential to mediate crystal contacts. The propensity of these mutants to form crystals in alternative screens in which the normal crystallization reservoir solutions were replaced with 1.5 M NaCl was also examined. The results were impressive: more than half of the mutants yielded a larger number of crystals with salt as the reservoir solution. This method greatly increased the variety of conditions that yielded crystals. Taken together, these results suggest a powerful crystallization strategy that combines surface engineering with efficient screening using standard and alternate reservoir solutions.

PubMed: 17452789
DOI: 10.1107/S0907444907010931

実験手法

X-RAY DIFFRACTION (1.88 Å)