Crystallization and preliminary crystallographic investigation of glycosomal pyruvate phosphate dikinase from Trypanosoma brucei

The PPi-dependent glycosomal enzyme pyruvate phosphate dikinase (PPDK) from Trypanosoma brucei is expressed in the insect stage of the parasite. Its p recise function there is still unclear, but the enzyme may catalyze the 're verse reaction' of transfer of phosphate from phosphoenolpyruvate (PEP) to generate pyruvate as a means of scavenging large amounts of pyrophosphate. This protein may represent a target for drug design against diseases caused by trypanosomes and related kinetoplastids. The recombinant protein is 918 amino acids long (predicted molecular mass similar or equal to 100 kDa and pI = 8.9). Crystallization conditions for the recombinant PPDK are reporte d that result in crystals that diffract X-rays to better than 3.0 Angstrom resolution. Their space group is P2(1)2(1)2, with unit-cell parameters a = 121.17, b = 153.5, c = 65.46 Angstrom, alpha = beta = gamma = 90 degrees. T he crystals, like the protein in solution, are sensitive to temperature and fail to diffract or diffract only to low resolution after ageing for two w eeks or longer.