Open AccessCrystallization and preliminary X‐ray diffraction analysis of the C‐terminal domain of the human spliceosomal DExD/H‐box protein hPrp22
Author(s) -
Kudlinzki Denis,
Nagel Christian,
Ficner Ralf
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109031844
Subject(s) - rna splicing , snrnp , spliceosome , rna , messenger rna , helicase , biology , crystallography , microbiology and biotechnology , chemistry , biophysics , biochemistry , gene
The Homo sapiens DExD/H‐box protein hPrp22 is a crucial component of the eukaryotic pre‐mRNA splicing machinery. Within the splicing cycle, it is involved in the ligation of exons and generation of the lariat and it additionally catalyzes the release of mature mRNA from the spliceosomal U5 snRNP. The yeast homologue of this protein, yPrp22, shows ATP‐dependent RNA‐helicase activity and is capable of unwinding RNA/RNA duplex molecules. A truncated construct coding for residues 950–1183 of human Prp22, comprising the structurally and functionally uncharacterized C‐terminal domain, was cloned into an Escherichia coli expression vector. The protein was subsequently overproduced, purified and crystallized. The crystals obtained diffracted to 2.1 Å resolution, belonged to the tetragonal space group P 4 1 2 1 2 or P 4 3 2 1 2, with unit‐cell parameters a = b = 78.2, c = 88.4 Å, and contained one molecule in the asymmetric unit.