Crystallization and preliminary X-ray crystallographic analysis of the unusual ferritin from Listeria innocua

1999 ◽  
Vol 55 (2) ◽  
pp. 552-553 ◽  
Author(s):  
Andrea Ilari ◽  
Carmelinda Savino ◽  
Simonetta Stefanini ◽  
Emilia Chiancone ◽  
Demetrius Tsernoglou
Keyword(s):  
Packing Density ◽  
Crystallographic Analysis ◽  
Listeria Innocua ◽  
Diffusion Method ◽  
Asymmetric Unit ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
Cell Dimensions ◽  
Vapour Diffusion ◽  
Unit Cell Dimensions

Single crystals of ferritin extracted from Listeria innocua have been obtained by the vapour-diffusion method using PEG 1000 as precipitant. The crystals are orthorhombic, space group P212121, with unit-cell dimensions a = 87.7, b = 137.5, c = 173.1 Å. The crystals diffract to 2.9 Å resolution on a rotating-anode X-ray source and to 2.35 Å resolution on a synchrotron X-ray source. The asymmetric unit contains one molecule formed by 12 subunits, corresponding to a packing density of 2.41 Å3 Da−1

scholarly journals Expression, purification, crystallization and preliminary X-ray crystallographic analysis of tomato β-galactosidase 4

2015 ◽  
Vol 71 (2) ◽  
pp. 153-156
Author(s):  
Masahiro Eda ◽  
Megumi Ishimaru ◽  
Toshiji Tada
Keyword(s):  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Galactosidase Activity ◽  
Asymmetric Unit ◽  
Orthorhombic Space Group ◽  
Solvent Content ◽  
X Ray ◽  
Signalling Molecules ◽  
Vapour Diffusion ◽  
Cell Wall Expansion

Plant β-galactosidases play important roles in carbohydrate-reserve mobilization, cell-wall expansion and degradation, and turnover of signalling molecules during ripening. Tomato β-galactosidase 4 (TBG4) not only has β-galactosidase activity but also has exo-β-(1,4)-galactanase activity, and prefers β-(1,4)-galactans longer than pentamers as its substrates; most other β-galactosidases only have the former activity. Recombinant TBG4 protein expressed in the yeastPichia pastoriswas crystallized by the sitting-drop vapour-diffusion method using PEG 10 000 as a precipitant. The crystals belonged to the orthorhombic space groupP212121, with unit-parametersa= 92.82,b= 96.30,c= 159.26 Å, and diffracted to 1.65 Å resolution. Calculation of the Matthews coefficient suggested the presence of two monomers per asymmetric unit (VM= 2.2 Å3 Da−1), with a solvent content of 45%.

Purification, crystallization and preliminary X-ray crystallographic analysis of recombinant murine Golgi mannosidase IA, a class I α-mannosidase involved in Asn-linked oligosaccharide maturation

1999 ◽  
Vol 55 (2) ◽  
pp. 571-573 ◽  
Author(s):  
Francois Vallée ◽  
Anita Lal ◽  
Kelley W. Moremen ◽  
P. Lynne Howell
Keyword(s):  
Transmembrane Domain ◽  
Crystallographic Analysis ◽  
Class I ◽  
Diffusion Method ◽  
Type Ii ◽  
Asymmetric Unit ◽  
Hanging Drop ◽  
X Ray ◽  
Cell Dimensions ◽  
Unit Cell Dimensions

Golgi mannosidase IA is a class I α-mannosidase which catalyzes the conversion of Man9GlcNAc2 or Man8GlcNAc2 oligosaccharide substrates to Man5GlcNAc2 during the maturation of Asn-linked oligosaccharides. The enzyme is a type II membrane protein, and a recombinant form of mannosidase IA from mouse, lacking the transmembrane domain, has been expressed in Pichia pastoris, purified to homogeneity and crystallized by the hanging-drop vapor-diffusion method. The crystals grow as thin rods, with unit-cell dimensions a = 54.9, b = 135.01, c = 69.9 Å. The crystals exhibit the symmetry of space group P2221 and diffract to 2.8 Å resolution. The asymmetric unit contains one monomer (∼53 kDa) and has a solvent content of 59%.

scholarly journals Cloning, purification and preliminary X-ray crystallographic analysis of the OmpA-like domain of peptidoglycan-associated lipoprotein fromAcinetobacter baumannii

2012 ◽  
Vol 68 (11) ◽  
pp. 1351-1353 ◽  
Author(s):  
Jung Hyun Song ◽  
Woo Cheol Lee ◽  
Jeong Soon Park ◽  
Seung Il Kim ◽  
Je Chul Lee ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Acinetobacter Baumannii ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Asymmetric Unit ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Vapour Diffusion ◽  
Strain Bl21

Peptidoglycan-associated lipoprotein (Pal) is one component of the Tol–Pal system that is involved in maintaining the integrity and stability of the outer membrane. The C-terminal OmpA-like domain of Pal interacts noncovalently with peptidoglycan. In this study, the OmpA-like domain of Pal fromAcinetobacter baumanniiwas overexpressed inEscherichia colistrain BL21 (DE3), purified and crystallized using the vapour-diffusion method. A native crystal diffracted to 1.4 Å resolution and belonged to space groupP61orP65, with unit-cell parametersa=b= 72.58,c= 44.65 Å, a calculated Matthews coefficient of 2.64 Å3 Da−1and one molecule per asymmetric unit.

scholarly journals Crystallization and preliminary X-ray analysis of the hypothetical deaminase RPB_0146 fromRhodopseudomonas palustrisHaA2

2014 ◽  
Vol 70 (11) ◽  
pp. 1560-1562
Author(s):  
Guofang Zhang ◽  
Dan Yu ◽  
Guodong Yang ◽  
Hui Dong ◽  
Tongcun Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rhodopseudomonas Palustris ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Hanging Drop ◽  
Unit Cell Parameters ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
Cell Parameters ◽  
Vapour Diffusion

RPB_0146, a putative deaminase fromRhodopseudomonas palustrisHaA2, was expressed inEscherichia coliBL21 (DE3) cells and purified using a His6tag by Ni2+-chelating affinity chromatography for X-ray crystallographic analysis. Diffraction-quality crystals were grown by the hanging-drop vapour-diffusion method at 289 K and diffracted to a resolution of 2.44 Å using a wavelength of 1.000 Å at the Photon Factory (KEK), Japan. The crystals belonged to the orthorhombic space groupP212121, with unit-cell parametersa= 66.26,b= 123.94,c= 155.95 Å.

scholarly journals Crystallization and preliminary X-ray crystallographic analysis of the XoGroEL chaperonin fromXanthomonas oryzaepv.oryzae

2014 ◽  
Vol 70 (5) ◽  
pp. 604-607 ◽  
Author(s):  
Huyen-Thi Tran ◽  
Tan-Viet Pham ◽  
Ho-Phuong-Thuy Ngo ◽  
Myoung-Ki Hong ◽  
Jeong-Gu Kim ◽  
...  
Keyword(s):  
Essential Role ◽  
Crystallographic Analysis ◽  
Xanthomonas Oryzae ◽  
Diffusion Method ◽  
Asymmetric Unit ◽  
Hanging Drop ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
Chaperonin Groel ◽  
Cellular Environment

Along with the co-chaperonin GroES, the chaperonin GroEL plays an essential role in enhancing protein folding or refolding and in protecting proteins against misfolding and aggregation in the cellular environment. TheXoGroELgene (XOO_4288) fromXanthomonas oryzaepv.oryzaewas cloned and the protein was expressed, purified and crystallized. The purified XoGroEL protein was crystallized using the hanging-drop vapour-diffusion method and a crystal diffracted to a resolution of 3.4 Å. The crystal belonged to the orthorhombic space groupP212121with 14 monomers in the asymmetric unit, with a correspondingVMof 2.7 Å3 Da−1and a solvent content of 54.5%.

Crystallization and preliminary X-ray analysis of β-amylase from Bacillus polymyxa

1999 ◽  
Vol 55 (4) ◽  
pp. 898-900
Author(s):  
Takashi Yamane ◽  
Hiroshi Tasaki ◽  
Fusako Matsumoto ◽  
Atsuo Suzuki ◽  
Nobuyuki Uozumi ◽  
...  
Keyword(s):  
Diffusion Method ◽  
Bacillus Polymyxa ◽  
Asymmetric Unit ◽  
Hanging Drop ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
Cell Dimensions ◽  
Protein Molecules ◽  
Density Map ◽  
Electron Density Map

A truncated β-amylase (E.C. 3.2.1.2) from Bacillus polymyxa has been crystallized using the hanging-drop vapour-diffusion method at 277 K. The crystals belong to the orthorhombic space group P212121 with cell dimensions a = 64.6, b = 141.9, c = 155.1 Å and diffract to 2.5 Å resolution. The asymmetric unit containing three protein molecules was derived from an electron-density map calculated at 4 Å resolution using MIR phases. This gives a Vm value of 2.36 Å3 Da−1.

Crystallization and preliminary X-ray studies of β-1,4-galactanase from Aspergillus aculeatus

1999 ◽  
Vol 55 (4) ◽  
pp. 929-930 ◽  
Author(s):  
C. Ryttersgaard ◽  
J.-C. N. Poulsen ◽  
S. Christgau ◽  
T. Sandal ◽  
H. Dalbøge ◽  
...  
Keyword(s):  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Aspergillus Aculeatus ◽  
Solvent Content ◽  
X Ray ◽  
Space Groups ◽  
Peg 400 ◽  
Cell Dimensions ◽  
Vapour Diffusion ◽  
Unit Cell Dimensions

Recombinant β-1,4-galactanase from Aspergillus aculeatus has been crystallized and characterized by X-ray diffraction. Crystals were obtained in hanging drops by vapour-diffusion under the conditions 30% PEG 400, 0.2 M CaCl2 and 0.1 M Na HEPES buffered to pH 7.5. The crystals diffract to 2.3 Å resolution and belong to one of the orthorhombic space groups I222 or I212121. The unit-cell dimensions are a = 60.42, b = 88.94 and c = 129.08 Å. With one molecule in the asymmetric unit, the corresponding solvent content is ∼48%.

Overproduction, purification, crystallization and preliminary X-ray diffraction studies of the human spliceosomal protein U5-15kD

1999 ◽  
Vol 55 (4) ◽  
pp. 888-890 ◽  
Author(s):  
Klaus Reuter ◽  
Ralf Ficner
Keyword(s):  
Escherichia Coli ◽  
Data Collection ◽  
Diffusion Method ◽  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
Gene Coding ◽  
Vapour Diffusion ◽  
Spliceosomal Protein

The gene coding for the human spliceosomal U5 snRNP-specific 15 kDa protein (U5-15kD) was overexpressed in Escherichia coli, its product purified to homogeneity and crystallized. Well diffracting single crystals were obtained by the vapour-diffusion method in hanging drops and subsequent macroseeding. The crystals belong to the orthorhombic space group P21212 with a = 62.3, b = 65.7, c = 37.1 Å. They diffract to at least 3.0 Å and contain one molecule in the asymmetric unit. A selenomethionine derivative of the protein was prepared and crystallized for multiwavelength anomalous diffraction (MAD) data collection.

Crystallization and preliminary X-ray diffraction studies of D-glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic archaeon Methanothermus fervidus

1999 ◽  
Vol 55 (7) ◽  
pp. 1353-1355 ◽  
Author(s):  
C. Charron ◽  
F. Talfournier ◽  
M. N. Isupov ◽  
G. Branlant ◽  
J. A. Littlechild ◽  
...  
Keyword(s):  
Diffusion Method ◽  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Hanging Drop ◽  
Data Set ◽  
Hyperthermophilic Archaeon ◽  
X Ray ◽  
Cell Dimensions ◽  
Methanothermus Fervidus ◽  
Unit Cell Dimensions

The homotetrameric holo-D-glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic archaeon Methanothermus fervidus has been crystallized in the presence of NADP+ using the hanging-drop vapour-diffusion method. Crystals grew from a solution containing 2-methyl-2,4-pentanediol and magnesium acetate. A native data set has been collected to 2.1 Å using synchrotron radiation and cryocooling. Diffraction data have been processed in the orthorhombic system (space group P21212) with unit-cell dimensions a = 136.7, b = 153.3, c = 74.9 Å and one tetramer per asymmetric unit.

Initiating a crystallographic study of UDP-galactopyranose mutase from Escherichia coli

1999 ◽  
Vol 55 (2) ◽  
pp. 399-402 ◽  
Author(s):  
Stephen A. McMahon ◽  
Gordon A. Leonard ◽  
Louise V. Buchanan ◽  
Marie-France Giraud ◽  
James H. Naismith
Keyword(s):  
Thin Plates ◽  
Diffusion Method ◽  
Data Sets ◽  
X Ray Diffraction ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
Crystallographic Study ◽  
Cell Dimensions ◽  
Unit Cell Dimensions ◽  
Platinum Derivative

UDP-galactopyranose mutase, the enzyme responsible for the conversion of UDP-galactopyranose to UDP-galactofuranose, has been crystallized in a form suitable for X-ray diffraction studies. UDP-galactofuranose is a key component of mycobacterial cell walls. Crystals of both the native protein and a selenomethionine variant have been grown by the vapour-diffusion method in hanging drops, and diffract to beyond 3.0 Å using synchrotron radiation. Equilibration was against a solution of 20%(w/v) polyethylene glycol (4K), 12%(v/v) 2-propanol, 0.1 M HEPES pH 7.6 at 293.5 K. Crystals grow as thin plates of dimensions 0.4 × 0.2 × ∼0.02 mm. They are orthorhombic, space group P21, with unit-cell dimensions a = 71.12, b = 58.42, c = 96.38 Å, β = 96.38°. 92% (native) and 94% (selenomethionine) complete data sets have been recorded to 2.9 Å (R merge = 5.0%) and 3.0 Å (R merge = 6.9%), respectively. The Matthews coefficient is 2.35 Å3 Da−1 for a dimer in the asymmetric unit, the solvent content being 47%. Diffraction data have also been recorded on a putative platinum derivative to 3.5 Å.

Sign in / Sign up

Export Citation Format

Share Document