Cloning, purification and crystallization of full-length human annexin 2.
Annexin 2, a Ca(2+)/phospholipid-binding protein, is involved in many biological processes, including membrane aggregation and the modulation of fibrinolytic activity. Here, the expression and purification of recombinant full-length human annexin 2 is reported, as well as crystals obtained by sitting-drop and hanging-drop vapor diffusion at 277 K. A condition consisting of 18% PEG 8000, 0.1 M sodium cacodylate pH 6.5, 0.2 M calcium acetate yielded long needles that diffracted to 3.20 A. Another condition, consisting of 2.5 M NaCl, 0.1 M acetate pH 4.5, 0.2 M Li(2)SO(4), gave crystals with unit-cell parameters a = 48.36, b = 62.86, c = 119.11 A that diffracted to 1.52 A. Both crystals belong to the orthorhombic P2(1)2(1)2(1) space group. The high-resolution 1.52 A data set was collected at ALS beamline 5.0.2 and is 93.0% complete, with an R(sym) of 4.5%. The structure of full-length annexin 2 will provide insight into how its N-terminal domain contributes to its functional role in a variety of biological processes.
D017306:Annexin A2; D017302:Annexins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray / Q000379:methods; D017931:DNA Primers; D006801:Humans; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
C472822:ANXA2 protein, human; D017306:Annexin A2; D017302:Annexins; D017931:DNA Primers; D011994:Recombinant Proteins