Expression, purification and preliminary X-ray crystallographic analysis of Arf1-GDP in complex with dimeric p23 peptide

摘要

Arf1 is a member of the Ras superfamily and is involved in COPI vesicle formation. Arf1-GDP can interact with dimeric p23. Here, human Arf1 (residues 18-181) was cloned, expressed and purified in Escherichia coli. For crystallization, Arf1-GDP was mixed with dimeric p23 peptide in a 1: 5 molar ratio. Crystals were obtained which diffracted to 2.7 angstrom resolution. The crystals belonged to space group P6(1)22, with unit-cell parameters a = b = 80.6, c = 336.0 angstrom, alpha = beta = 90, gamma = 120 degrees. The asymmetric unit of the crystals contained two molecules, with a Matthews coefficient of 3.2 angstrom(3) Da(-1) and a solvent content of 61.9%.