Crystallization and preliminary X-ray analysis of Sau3AI/E64A mutant protein

Chunyan Xu; Jiaping Song; Yu Ding; Feng Yu; Lihua Sun; Lin Tang; Xiaojian Hu; Zhihong Zhang; Jianhua He

doi:10.2174/092986607780782812

Crystallization and preliminary X-ray analysis of Sau3AI/E64A mutant protein

Protein Pept Lett. 2007;14(5):505-6. doi: 10.2174/092986607780782812.

Authors

Chunyan Xu¹, Jiaping Song, Yu Ding, Feng Yu, Lihua Sun, Lin Tang, Xiaojian Hu, Zhihong Zhang, Jianhua He

Affiliation

¹ Shanghai Institutes of Applied Physics, Chinese Academy of Sciences, 201800 Shanghai PR China.

PMID: 17584179
DOI: 10.2174/092986607780782812

Abstract

Sau3AI is a type II restriction endonuclease that recognizes the palindromic sequence 5'-GATC-3' and cleaves 5' to G residue on each strand. The E64A mutant full length protein was cloned and expressed in Escherichia coli. The purified (His) (6)-tagged protein has monomer and dimer fraction and was crystallized by the hanging-drop vapor-diffusion technique. The dimer protein crystals can diffract to 3.0A. resolution and the monomer protein crystals can diffract to better than 2.8A. resolution. One completed dataset has been collected and it shows that the monomer orthorhombic Sau3AI/E64A crystal is in space group C2221 with unit cell parameters (69.44, 197.60, 191.46, 90, 90, 90) and contains two molecules in one asymmetric unit.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Substitution
Cloning, Molecular
Crystallization
Crystallography, X-Ray
Deoxyribonucleases, Type II Site-Specific / chemistry*
Deoxyribonucleases, Type II Site-Specific / genetics
Escherichia coli / metabolism
Staphylococcus aureus / enzymology

Substances

Deoxyribonucleases, Type II Site-Specific
GATC-specific type II deoxyribonucleases