Crystallization and preliminary X-ray analysis of the cytoplasmic domain of human erythrocyte band 3.
A cytoplasmic domain of the human erythrocyte membrane protein band 3 (M(r) = 42,500), residues 1-379, expressed in and purified from E. coli, has been crystallized by the method of vapor diffusion in sitting drops with subsequent streak-seeding at room temperature. Initial crystals were grown from solutions containing 65-68% saturated ammonium sulfate at pH 4.9 and 2 mg/ml protein. Subsequent streak-seeding into solutions of 50-53% ammonium sulfate at pH 4.9 and 7 mg/ml protein produced single crystals suitable for X-ray analysis, which contained pure protein as revealed by gel electrophoresis. The crystals belong to the monoclinic space group C2 with cell dimensions of a = 178.8 A, b = 90.5 A, c = 122.1 A, and beta = 131.3 degrees and diffract at least to 2.7 A resolution (at 100 K). A self-rotation function shows the presence of approximate 222 local symmetry.
D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D003593:Cytoplasm / Q000737:chemistry; D004910:Erythrocyte Membrane / Q000737:chemistry*; D006801:Humans; D011487:Protein Conformation
D001457:Anion Exchange Protein 1, Erythrocyte