Definition of domain boundaries and crystallization of the SMN Tudor domain.
Spinal muscular atropy (SMA) is the major genetic disease leading to childhood mortality and is caused by mutations in or deletions of the smn1 gene. The human survival of motor neurons (SMN) protein encoded by this gene plays an important role in the assembly of snRNPs (small nuclear ribonucleoprotein complexes) via binding to the spliceosomal Sm proteins. The tails of these Sm proteins contain symmetrically dimethylated arginines that are recognized by the central SMN Tudor domain. To gain insight in the molecular basis of this specific interaction, the SMN Tudor domain has been crystallized. The rapid crystallization of the protein and the high stability of the crystals is facilitated by redefinition of domain boundaries based on NMR relaxation experiments and the previously determined solution structure. The crystals diffract to high resolution (1.8 A) and a complete data set has been collected from a hexagonal crystal form (P6(1)/P6(5)), with unit-cell parameters a = b = 27.65, c = 110.30 A, alpha = beta = 90, gamma = 120 degrees. Crystal soaks and co-crystallization with peptides derived from the Sm protein tails have been initiated. Molecular replacement with the NMR coordinates is under way.
Sprangers
Remco
R
Structural and Computational Biology, EMBL, Meyerhofstrasse 1, D-69012, Heidelberg, Germany.
Selenko
Philipp
P
Sattler
Michael
M
Sinning
Irmgard
I
Groves
Matthew R
MR
D003460:Crystallization; D018360:Crystallography, X-Ray; D017362:Cyclic AMP Response Element-Binding Protein; D006801:Humans; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins; D055532:SMN Complex Proteins; D055533:Survival of Motor Neuron 1 Protein
D017362:Cyclic AMP Response Element-Binding Protein; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; D055532:SMN Complex Proteins; C526471:SMN1 protein, human; D055533:Survival of Motor Neuron 1 Protein