Cloning, expression, purification, crystallization and preliminary crystallographic analysis of gamma-filamin repeat 23.
Human gamma-filamin is a protein of 2705 amino-acid residues that localizes mainly in the myofibrillar Z-disc and to smaller extent in the subsarcolemmal region of striated muscle cells. gamma-Filamin consists of an N-terminal actin-binding domain followed by a long rod-shaped region. The rod-shaped region consists of 24 immunoglobulin-like domains that form a platform for interaction with different transmembrane, cell-signalling and cytoskeletal proteins. gamma-Filamin repeat 23 was indicated as being necessary for binding to the muscle-specific subsarcolemmal proteins gamma- and delta-sarcoglycan and the myofibrillar protein FATZ1. The recombinant gamma-filamin repeat 23 was crystallized using the hanging-drop vapour-diffusion method, which yielded needle-shaped diffraction-quality crystals. Diffraction data were collected to 2.05 angstroms resolution using 1.2 angstroms wavelength synchrotron radiation. Preliminary structural analysis shows one molecule, with predominantly beta secondary-structure elements, per asymmetric unit.
Sjekloća
Ljiljana
L
Structural Biology Laboratory, Elettra - Sincrotrone Trieste in Area Science Park, S.S. 14 Km 163.5 loc. Basovizza, 34012 Trieste, Italy.
Sjöblom
Björn
B
Polentarutti
Maurizio
M
Djinović Carugo
Kristina
K
D000199:Actins / Q000737:chemistry; D003001:Cloning, Molecular; D003285:Contractile Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D064448:Filamins; D006639:Histidine / Q000737:chemistry; D006801:Humans; D008840:Microfilament Proteins / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D013917:Thrombin / Q000737:chemistry; D014961:X-Ray Diffraction
D000199:Actins; D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D011994:Recombinant Proteins; D006639:Histidine; D013917:Thrombin