Open AccessCloning, overexpression, purification, crystallization and preliminary X‐ray diffraction analysis of glyceraldehyde‐3‐phosphate dehydrogenase from Antheraea mylitta
Author(s) -
Mukherjee Somnath,
Maity Samita,
Roy Sobhan,
Ghorai Suvankar,
Chakrabarti Mrinmay,
Agarwal Rachit,
Dutta Debajyoti,
Ghosh Ananta Kumar,
Das Amit Kumar
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910903214x
Subject(s) - orthorhombic crystal system , crystallization , dehydrogenase , crystallography , escherichia coli , glyceraldehyde 3 phosphate dehydrogenase , chemistry , microbiology and biotechnology , diffraction , biology , biochemistry , crystal structure , enzyme , physics , gene , optics , organic chemistry
Glyceraldehyde‐3‐phosphate dehydrogenase from Antheraea mylitta (AmGAPDH) was cloned in pQE30 vector, overexpressed in Escherichia coli M15 (pREP4) cells and purified to homogeneity. The protein was crystallized using the hanging‐drop vapour‐diffusion method. The crystals belonged to the orthorhombic space group I 222, with unit‐cell parameters a = 85.81, b = 133.72, c = 220.37 Å. X‐ray diffraction data were collected and processed to a maximum resolution of 2.2 Å. The presence of three molecules in the asymmetric unit gave a Matthews coefficient ( V M ) of 2.80 Å 3 Da −1 , with a solvent content of 56.08%.