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Cloning, overexpression, purification, crystallization and preliminary X-ray diffraction analysis of glyceraldehyde-3-phosphate dehydrogenase from Antheraea mylitta.

ABSTRACT: 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%.

SUBMITTER: Mukherjee S

PROVIDER: S-EPMC2795606 | biostudies-literature | 2009 Sep

REPOSITORIES: biostudies-literature

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Cloning, overexpression, purification, crystallization and preliminary X-ray diffraction analysis of glyceraldehyde-3-phosphate dehydrogenase from Antheraea mylitta.

Mukherjee Somnath S 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

Acta crystallographica. Section F, Structural biology and crystallization communications 20090826 Pt 9

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 ...[more]

PMID: 19724138

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