Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Enpp6

Enpp (ectonucleotide phosphodiesterase/pyrophosphatase) 6 is a membrane-bound glycoprotein that hydrolyzes choline-containing compounds such as lysophosphatidylcholine and glycerophosphorylcholine, and presumably participates in choline metabolism. The catalytic domain of mouse Enpp6 was expressed in HEK293T cells, purified using the TARGET tag/P20.1-Sepharose system and crystallized. An X-ray diffraction data set was collected to 1.8 Å resolution. The crystal belonged to space group P1, with unit-cell parameters a = 63.7, b = 68.8, c = 69.7 Å, α = 60.6, β = 87.0, γ = 68.1°. Assuming the presence of two protein molecules per asymmetric unit, the solvent content was estimated to be 49.5%.

Source: Acta Crystallographica Section F - May 24, 2014 Category: Biochemistry Authors: Morita, J.Kato, K.Mihara, E.Ishitani, R.Takagi, J.Nishimasu, H.Aoki, J.Nureki, O. Tags: Enpp6 choline N-glycosylation crystallization communications Source Type: research