Purification, crystallization, and preliminary X-ray diffraction study of purine nucleoside phosphorylase from E. coli

Abstract

Crystals of E. coli purine nucleoside phosphorylase were grown in microgravity by the capillary counter-diffusion method through a gel layer. The X-ray diffraction data set suitable for the determination of the three-dimensional structure at atomic resolution was collected from one crystal at the Spring-8 synchrotron facility to 0.99 A resolution. The crystals belong to sp. gr. P21 and have the following unit-cell parameters: a = 74.1 A, b = 110.2 A, c = 88.2 A, α = γ = 90°, β = 111.08°. The crystal contains six subunits of the enzyme comprising a hexamer per asymmetric unit. The hexamer is the biological active form of E. coli. purine nucleoside phosphorylase.