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The nuclear import of spliceosomal UsnRNPs is mediated by the transport adaptor snurportin 1 (SPN1), which specifically recognizes the 2,2,7-trimethylguanosine (m3G) cap at the 5' end of UsnRNAs. Human SPNI was overexpressed as a OST-fusion protein in Escherichia coli and purified to homogeneity. Since full-length SPNI did not crystallize, limited proteolysis experiments were performed and stable digestion products were analyzed for functionality with respect to m3G cap-binding activity and subsequently used for crystallization trials. Well diffracting single crystals of a truncated SPNI m30 cap-binding domain (residues 79-300) were obtained after two rounds of seeding. The crystals belong to space group P41212 or P43212, with unit-cell parameters a = b = 57.47, c = 130.09 Å, α = β = γ = 90°. Crystals contain one molecule in the asymmetric unit and diffract to a resolution limit of 2.9 Å. [ABSTRACT FROM AUTHOR]

The nuclear import of spliceosomal UsnRNPs is mediated by the transport adaptor snurportin 1 (SPN1), which specifically recognizes the 2,2,7-trimethylguanosine (m(3)G) cap at the 5' end of UsnRNAs. Human SPN1 was overexpressed as a GST-fusion protein in Escherichia coli and purified to homogeneity. Since full-length SPN1 did not crystallize, limited proteolysis experiments were performed and stable digestion products were analyzed for functionality with respect to m(3)G cap-binding activity and subsequently used for crystallization trials. Well diffracting single crystals of a truncated SPN1 m(3)G cap-binding domain (residues 79-300) were obtained after two rounds of seeding. The crystals belong to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 57.47, c = 130.09 A, alpha = beta = gamma = 90 degrees. Crystals contain one molecule in the asymmetric unit and diffract to a resolution limit of 2.9 A.