Crystallization of the actin-binding domain of human alpha-actinin: analysis of microcrystals of SeMet-labelled protein.
Alpha-actinin forms antiparallel homodimers that cross-link actin filaments from adjacent sarcomeres within the Z-discs of striated muscle. The N-terminal actin-binding domain (ABD) is composed of two calponin homology (CH) domains followed by four spectrin-like repeats and a calmodulin-like EF-hand domain at the C-terminus. The ABD of human alpha-actinin crystallizes in space group P2(1), with unit-cell parameters a = 101.9, b = 38.4, c = 154.9 A, beta = 109.2 degrees. A complete native data set from a native crystal was collected extending to 2.0 A resolution and a single-wavelength anomalous dispersion (SAD) data set to 2.9 A resolution was collected from a selenomethionine-labelled microcrystal using the microfocusing beamline ID-13 at the ESRF. Analysis of the anomalous contribution shows a rapid decrease in the sigma(normal)/sigma(anomal) ratio owing to radiation damage.
Ekström
Fredrik
F
Umeå Center for Molecular Pathogenisis, Umeå Universit, Sweden.
Stier
Gunter
G
Sauer
Uwe H
UH
D000185:Actinin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D000199:Actins / Q000378:metabolism*; D003001:Cloning, Molecular; D003460:Crystallization; D006801:Humans; D007202:Indicators and Reagents; D011485:Protein Binding; D012645:Selenomethionine / Q000737:chemistry
D000199:Actins; D007202:Indicators and Reagents; D000185:Actinin; D012645:Selenomethionine