Cloning, overexpression, purification, crystallization and preliminary X-ray diffraction analysis of glyceraldehyde-3-phosphate dehydrogenase from Antheraea mylitta.
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 I222, with unit-cell parameters a = 85.81, b = 133.72, c = 220.37 A. X-ray diffraction data were collected and processed to a maximum resolution of 2.2 A. The presence of three molecules in the asymmetric unit gave a Matthews coefficient (V(M)) of 2.80 A(3) Da(-1), with a solvent content of 56.08%.
Mukherjee
Somnath
S
Department of Biotechnology, Indian Institute of Technology, Kharagpur 721302, India.
Maity
Samita
S
Roy
Sobhan
S
Ghorai
Suvankar
S
Chakrabarti
Mrinmay
M
Agarwal
Rachit
R
Dutta
Debajyoti
D
Ghosh
Ananta Kumar
AK
Das
Amit Kumar
AK
D000818:Animals; D012831:Bombyx / Q000201:enzymology*; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases