Improving the diffraction of full-length human selenomethionyl metavinculin crystals by streak-seeding.
Metavinculin is an alternatively spliced isoform of vinculin that has a 68-residue insert in its tail domain (1134 total residues) and is exclusively expressed in cardiac and smooth muscle tissue, where it plays important roles in myocyte adhesion complexes. Mutations in the metavinculin-specific insert are associated with dilated cardiomyopathy (DCM) in man. Crystals of a DCM-associated mutant of full-length selenomethionine-labeled metavinculin grown by hanging-drop vapor diffusion diffracted poorly and were highly sensitive to radiation, preventing the collection of a complete X-ray diffraction data set at the highest possible resolution. Streak-seeding markedly improved the stability, crystal-growth rate and diffraction quality of DCM-associated mutant metavinculin crystals, allowing complete data collection to 3.9 Å resolution. These crystals belonged to space group P4(3)2(1)2, with two molecules in the asymmetric unit and unit-cell parameters a=b=170, c=211 Å, α=β=γ=90°.
Cell Adhesion Laboratory, Department of Cancer Biology, The Scripps Research Institute, Scripps Florida, Jupiter, FL 33458, USA.
Erumbi S
ES
Rangarajan
Tina
T
Izard
D002311:Cardiomyopathy, Dilated / Q000235:genetics; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D050505:Mutant Proteins / Q000737:chemistry; D012645:Selenomethionine / Q000737:chemistry*; D016596:Vinculin / Q000737:chemistry*; D014961:X-Ray Diffraction / Q000379:methods*
D050505:Mutant Proteins; C103334:metavinculin; D016596:Vinculin; D012645:Selenomethionine