purified native isozyme alphaCA1, 0.002 ml of 20 mg/ml protein in 25 mM Tris-HCl, pH 7.5, is mixed with 0.002 ml of reservoir solution, containing 1.0 M ammonium sulfate and 100 mM Tris-HCl, pH 8.5, and equilibrated against 0.7 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 1.88 A resolution, MAD method
purified recombinant detagged enzyme CrCAH3 with inhibitors phosphate and acetazolamide, hanging drop vapor diffusion method, mixing of 0.001 ml of 3.6 mg/l protein in mM Tris-HCl, pH 8.0, and 150 mM NaCl, and 1 mM ligand, with 0.001 ml of reservoir solution containing 2.5 M NH4H2PO4 and 0.1 M Tris-HCl, pH 8.0, to a final pH of pH 4.1, and equilibration over 1 ml reservoir solution, at 18°C, method optimization, X-ray diffraction structure determination and analysis at 2.6-2.7 A resolution
crystallized by the hanging drop vapour-diffusion method, monoclinic crystal system, crystals belong to space group P2(1), with unit-cell parameters a : 47.0 A, b : 119.9 A, c : 58.5 A
2 different tetragonal crystal forms, form 1 space group P4(2)2(1)2, unit cell dimensions a : b : 68.54 A, c : 85.88A, form 2 space group P4(3)22, a : : b : 81.24 A, c : 162.14 A
CynT2, form 1, space group P4(2)212, unit-cell parameters a : b : 68.54 A, c : 85.88 A, form 2, space group P4(3)22, a : b : 81.00 A, c : 161.98 A, form 3, space group P2(1), a : 48.21, b : 140.73, c : 72.57, selenomethionine-substituted crystals, space group P4(3)22, a : b : 81.24 A, c : 162.14 A
purified recombinant Co-HICA by hanging drop vapor diffusion, 10 mg/ml protein crystallized in 0.2 M sodium acetate, 0.1 M Tris-HCl, pH 8.5, 0.1 M (NH4)2SO4, and 27% PEG 4000, 22°C, several days, X-ray diffraction structure determination and analysis at 2.5 A resolution
purified wild-type and mutant enzymes, 12 mg/ml protein mixed with 0.7 M sodium potassium tartrate, 0.10 M HEPES, pH 7.5, for tetragonal crystals and with 1.8 M ammonium sulfate, 4% PEG 400, 0.10 M HEPES, pH 7.5, for monoclinic crystals, 22°C, 2-3 days, crystals are soaked for 1-2 min in artificial mother liquor plus either 30% glucose or 30% glucose and 100 mM NaHCO3, X-ray diffraction structure determination and analysis
all variants crystallized in the orthorhombic P2(1) 2(1) 2 (1) space group and are highly isomorphous with approximate unit cell dimensions a=42, b=72 and c=75 A and diffract between 1.56 and 2.0 A resolution
carbonic anhydrase II complexed with two aromatic sulfonamide inhibitors N-(4-sulfamoylphenyl)-2-(thiophen-2-yl)acetamide and N-(2-fluoro-4-sulfamoylphenyl)-2-(thiophen-2-yl)acetamide incorporating 2-thienylacetamido moieties, hanging drop vapor diffusion method, drops of 0.005 ml containing 0.5 mM hCA II, 1 mM inhibitor, 0.1% DMSO, 0.8 M sodium citrate, 50 mM Tris-HCl, pH 8.0,are equilibrated against 1 ml precipitant solution containing 1.6 M sodium citrate and 50 mM Tris-HCl, pH 8.0, at room temperature, X-ray diffraction structure determination and analysis at 1.6-1.7 A resolution
crystal structures of C-terminal hexahistidine-tagged carbonic anhydrase III and F198L carbonic anhydrase III, hanging-drop vapour diffusion method, 2.1 A resolution
crystals obtained by hanging drop technique, hCA II-sulfamate, space group P2(1), a : 42.32 A, b : 41.48 A, c : 72.44 A, hCA II-sulfamide, space group P2(1), a : 42.70 A, b : 41.70 A, c : 73.00 A
crystals obtained using the hanging drop vapor diffusion method, isozyme CA I Michigan I, space group P2(1)2(1)2(1), with unit cell dimensions parameters a : 62.50 A, b : 72.13 A, c : 121.54 A, isozyme CA I Michigan 1-Zn(II)2, space group P2(1)2(1)2(1), with unit cell dimensions parameters a : 62.68 A, b : 71.36 A, c : 120.91A
crystals of K64H, R67H, and K64H/R67N HCA III are grown by hanging drop vapor diffusion method. X-ray crystal structures of site-specific mutants of human carbonic anhydrase III (HCA III): K64H, R67H, and K64H/R67N HCA III
crystals of the HCA II single-site mutants Y7F, N62L, and N67L are obtained using the hanging-drop vapor diffusion method. All crystals are isomorphous and belong to space group P2(1) with mean unit cell dimensions: a = 42.7 A, b = 41.6 A, c = 72.9 A and beta = 104.6°
in complex with inhibitor RWJ-37497, crystals obtained by the hanging-drop vapour-diffusion method, space group P2(1) with unit cell parameters a : 42.28 A, b : 41.39 A, c : 72.51 A
isoform CA II in complex with 6,7-dimethoxy-3,4-dihydroisoquinoline-2(1H)-sulfonamide, hanging drop vapor diffusion method, using 2.5 M (NH4)2SO4, 0.3 M NaCl, 100 mM Tris-Cl, pH 8.2
isozyme CA XIII in the unbound state and in complex with acetazolamide, hanging drop vapour diffusion method, in 20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM dithiothreitol with 30% (w/v) polyethylene glycol 4000, 0.2 M ammonium acetate, 0.1 M sodium acetate, pH 4.6
purified enzyme in complex with inhibitors (S)-4-(4-acetyl-3-benzylpiperazine-1-carbonyl)benzene-1-sulfonamide and (S)-4-[2-benzyl-4-(4-sulfamoylbenzoyl)piperazine-1-carbonyl]benzene-1-sulfonamide, sitting drop vapor diffusion method, mixing of 0.002 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 9.0, with 0.002 ml of a solution of 28-31% PEG 4000, 0.2 M sodium acetate, 0.1 M Tris, pH 8.5-9.0 and equilibration against the same solution at 23°C, fifteen days, afterwards the crystals are soaked in 5 mM inhibitor solutions for 3 days, X-ray diffraction structure determination and analysis at 1.5-1.6 A resolution
purified isozyme CA II (chimeric CA IX mimic) mutant in complex with inhibitors 4-phenylbenzene-1-sulfonamide, 4-(2-methylphenyl)benzene-1-sulfonamide, 4-(3-formylphenyl)benzene-1-sulfonamide, and 4-(3-quinolinyl)benzene-1-sulfonamide, sitting drop vapor diffusion method, mixing of 300-500 nl of 20 mg/mL of CA II and 41 mg/ml of the CA IX mimic solution with crystallization solution containing 1.6 M sodium citrate, 50 mM Tris-HCl, pH 7.8, in a ratio of 3:5 or 1:1, 25°C, 1 week, X-ray diffraction structure determination and analysis at 1.55-1.69 A resolution (PDB IDS 5SZ0, 5SZ1, 5SZ2, and 5SZ3), molecular replacement using the enzyme structure PBD ID 3KS3 without zinc and solvent as search model
purified isozyme CA IX in complex with inhibitors 4-phenylbenzene-1-sulfonamide, 4-(2-methylphenyl)benzene-1-sulfonamide, 4-(3-formylphenyl)benzene-1-sulfonamide, and 4-(3-quinolinyl)benzene-1-sulfonamide, X-ray diffraction structure determination and analysis at 1.15-1.78 A resolution (PDB IDS 5SZ4, 5SZ5, 5SZ6, and 5SZ7), molecular replacement using the enzyme structure PBD ID 3KS3 without zinc and solvent as search model
purified recombinant differently isotope-labeled isozyme hCAII, 14 mg/ml protein, both vapour diffusion and microbatch under-oil techniques are valuated, hanging and sitting drop set-ups with 0.01 ml drops, microseeding, deuteration of hCA II does not affect the crystal structure. Large, single crystals grow for both H and D enzyme versions in microbatch under-oil experiments at pH 8.5, room temperature, X-ray diffraction structure determination and analysis
purified recombinant differently isotope-labeled isozyme hCAIX, 17 mg/ml protein, both vapour diffusion and microbatch under-oil techniques are valuated, hanging and sitting drop set-ups with 0.01 ml drops, microseeding, room temperature, deuteration of hCA XI does not affect the crystal structure, but for D-hCA IX mimic no crystals appear in any of the under-oil batch trays at all pHs tested, while H-hCA IX SV gives several good crystals per drop at pH 8.5. X-ray diffraction structure determination and analysis
purified recombinant HCA II, crystals are prepared at pH 6.0, pH 8.5, and pH 11.0, hanging drop vapor diffusion method, crystal preparation from apo-Co(II)-HCA II by mixing 0.005 ml of protein solution with 0.005 ml of precipitant solution, containing 1.4 M sodium citrate, and 100 mM Tris-HCl, pH 9.0, at room temperature, and soaking of the crystals in 100 mM CoCl2, 1.4 M sodium citrate, 50 mM Tris-Cl, 50 mM 3-(cyclohexylamino)propanesulfonic acid with pH values of 6.0, 8.5 and 11.0, respectively, 2-3days, X-ray diffraction structure determination and analysis at 1.5-1.6 A resolution
purified recombinant human CA II in complex with imidazole-based activator CAA12i, hanging drop vapor diffusion method, mixing of 10 mg/ml protein and 0.2 mM ligand in 50 mM Tris-HCl, pH 7.8, with reservoir solution, containing 50 mM Tris HCl and 1.3 M sodium citrate, pH 7.4, in a 1:1 ratio, 23°C, 7 days without agitation, X-ray diffraction structure determination and analysis at 1.6 A resolution
purified recombinant K170 HCA II variants, hanging drop vapour diffusion method, 2-3days, X-ray diffraction structure determination at room temperature and analysis
the hCA IIN-hydroxysulfamide complex is cocrystallized at 4°C by the hanging drop vapor diffusion method. X-ray crystal structure of the human hCA II-N-hydroxysulfamide adduct
purified recombinant enzyme, sitting drop vapour diffusion, using a precipitant solution consisting of 2% v/v Tacsimate, pH 4.0, 0.1 M sodium acetate trihydrate, pH 4.6, and 16% w/v PEG 3350, 10 days at 17°C, X-ray diffraction structure determination and analysis at 2.6 A resolution
cubic crystals, space group P2(1)3, Zn-Cam, unit cell a : b : c: 82.64 A, Zn-Cam-HCO3-, unit cell a : b : c: 82.51 A, Zn-Cam-SO42-, unit cell a : b : c: 82.68 A, Co-Cam, unit cell a : b : c: 82.34 A, Co-Cam-HCO3-, unit cell a : b : c: 82.58 A, Co-Cam-SO42-, unit cell a : b : c: 82.61 A
the crystal structure of the enzyme is determined at 2.8 A resolution using the method of multiple isomorphous replacement in combination with 3fold non-crystallographic symmetry averaging and phase extension
in complex with Ca2+, sitting drop vapour diffusion method, with 22.5% PEG 4000 in MES-NaOH pH 5.8 and 0.1 M lithium chloride as the precipitating agent
Nce103 in complex with a substrate analog at 2.04 A resolution, hanging drop vapor diffusion method, using 20% (w/v) PEG4000, 0.1 M sodium citrate, pH 5.6, 0.1 M sodium acetate, 20% (v/v) ethylene glycol, at 16°C
purified recombinant His-tagged enzyme incomplex with inhibitor acetazolamide, hanging drop vapor diffusion method, mixing of 0.001 ml of 8 mg/ml protein in 20 mM Tris, pH 8.3, and a 10-molar excess of the inhibitor with 0.001 m of reservoir solution containing 30% w/v PEG monomethyl ether 550, 0.05 M MgCl2, and 0.1 M HEPES, pH 7.5, and equilibration against 0.5 ml of reservoir solution, 1 week, room temperature, X-ray diffraction structure determination and analysis at 1.95 A resolution, molecular replacement using the structure of enzyme SspCA from Sulfurihydrogenibium yellowstonensis (PDB ID 4G7A) as a search model
purified recombinant truncated enzyme CcaA220, by sitting drop vapor diffusion method, mixing of 0.001 ml of 7 mg/ml protein solution with 0.001 ml of well solution containing 2.0 M Na formate and 0.1 M Na acetate, pH 4.6, X-ray diffraction structure determination and analysis at 1.45 A resolution, molecular replacement using the structure of a catalytic dimer (residues 121-318) from Pisum sativum beta-CA (PDB ID 1ekj) as a search model