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

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Sep 1;65(Pt 9):937-40. doi: 10.1107/S174430910903214X. Epub 2009 Aug 26.

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%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bombyx / enzymology*
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Glyceraldehyde-3-Phosphate Dehydrogenases / chemistry*
  • Glyceraldehyde-3-Phosphate Dehydrogenases / isolation & purification*
  • Recombinant Proteins / chemistry
  • X-Ray Diffraction*

Substances

  • Recombinant Proteins
  • Glyceraldehyde-3-Phosphate Dehydrogenases