scholarly journals Crystallization and preliminary X-ray crystallographic studies of the coiled-coil domain of PIST

2013 ◽  
Vol 69 (4) ◽  
pp. 468-471
Author(s):  
Young-Cheul Shin ◽  
Eun Kyoung Seo ◽  
Ju-Hong Jeon ◽  
Hyun Ho Park
Keyword(s):  
Escherichia Coli ◽  
Membrane Trafficking ◽  
Coiled Coil ◽  
Cancer Development ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Coiled Coil Domain ◽  
Discs Large

PIST [PDZ (PSD-95, Discs-large and ZO-1) protein interacting specifically with TC10] functions as a regulator of membrane trafficking with Rab6A. Recently, the involvement of the fusion of PIST with ROS1 in cancer development has been identified. In this study, the coiled-coil domain of PIST, which is the domain responsible for interaction with Rab6A and fusion with ROS1, corresponding to amino acids 29–133, was overexpressed inEscherichia coliusing engineered C-terminal His tags. The coiled-coil domain of PIST was then purified to homogeneity and crystallized at 293 K. Finally, X-ray diffraction data were collected to a resolution of 4.0 Å from a crystal belonging to the hexagonal space groupP6222 orP6422, with unit-cell parametersa=b= 85.19,c= 240.09 Å, γ = 120.00°.

Preparation and preliminary X-ray diffraction studies of a new crystal form of L-asparaginase from Escherichia coli

1999 ◽  
Vol 55 (9) ◽  
pp. 1616-1617
Author(s):  
I. Polikarpov ◽  
R. T. de Oliveira ◽  
J. Abrahão-Neto
Keyword(s):  
Escherichia Coli ◽  
Synchrotron Radiation ◽  
Crystal Form ◽  
Asymmetric Unit ◽  
Chemotherapeutic Drug ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Synchrotron Radiation Diffraction

L-Asparaginase is an enzyme which hydrolyzes asparagine to produce aspartic acid and ammonia. It is an effective chemotherapeutic drug, especially in the treatment of acute lymphoblastic leukaemia in children. The enzyme from Escherichia coli was crystallized in a new crystal form with space group C2, unit-cell parameters a = 76.3 (0), b = 134.6 (2), c = 64.8 (7) Å, β = 110.5 (1)° and a dimer in the asymmetric unit. Synchrotron-radiation diffraction data have been collected to 1.95 Å resolution.

scholarly journals Preliminary X-ray crystallographic studies of the TIR domain of human Toll-like receptor 6

2014 ◽  
Vol 70 (8) ◽  
pp. 1053-1055 ◽  
Author(s):  
Tae-ho Jang ◽  
Hyun Ho Park
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Immune Response ◽  
Innate Immune ◽  
Toll Like Receptor ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Tir Domain

Toll-like receptor (TLR) proteins have been identified and shown to play a role in the innate immune response. TLR6 associated with TLR2 can recognize diacylated lipoprotein. In this study, the human TLR6 TIR domain corresponding to amino acids 640–796 was overexpressed inEscherichia coliusing engineered C-terminal His tags. The TLR6 TIR domain was then purified to homogeneity and crystallized at 20°C. Finally, X-ray diffraction data were collected to a resolution of 2.2 Å from a crystal belonging to space groupC2, with unit-cell parametersa= 127.60,b= 44.20,c= 75.72 Å, β = 118.89°

scholarly journals Crystallization and X-ray diffraction studies of a two-domain laccase fromStreptomyces griseoflavus

2015 ◽  
Vol 71 (9) ◽  
pp. 1200-1204 ◽  
Author(s):  
Svetlana Tishchenko ◽  
Azat Gabdulkhakov ◽  
Liubov Trubitsina ◽  
Alexander Lisov ◽  
Marina Zakharova ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Phenolic Compounds ◽  
Synchrotron Radiation ◽  
Monoclinic Space Group ◽  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters

Laccase (EC 1.10.3.2) is one of the most common copper-containing oxidases; it is found in many organisms and catalyzes the oxidation of primarily phenolic compounds by oxygen. Two-domain laccases have unusual thermostability, resistance to inhibitors and an alkaline optimum of activity. The causes of these properties in two-domain laccases are poorly understood. A recombinant two-domain laccase (SgfSL) was cloned from the genome ofStreptomyces griseoflavusAc-993, expressed inEscherichia coliand purified to homogeneity. The crystals of SgfSL belonged to the monoclinic space groupP21, with unit-cell parametersa= 74.64,b= 94.72,c= 117.40 Å, β = 90.672°, and diffraction data were collected to 2.0 Å resolution using a synchrotron-radiation source. Two functional trimers per asymmetric unit correspond to a Matthews coefficient of 1.99 Å3 Da−1according to the monomer molecular weight of 35.6 kDa.

scholarly journals Complexation of the nickel and cobalt transcriptional regulator RcnR with DNA

2020 ◽  
Vol 76 (1) ◽  
pp. 25-30
Author(s):  
Chao Li ◽  
Joseph W. Vavra ◽  
Carolyn E. Carr ◽  
Hsin-Ting Huang ◽  
Michael J. Maroney ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Transcription Factor ◽  
Transcriptional Regulator ◽  
Bacterial Cells ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Dna Binding Site ◽  
Cobalt And Nickel

RcnR is a transcription factor that regulates the homeostasis of cobalt and nickel in bacterial cells. Escherichia coli RcnR was crystallized with DNA that encompasses the DNA-binding site. X-ray diffraction data were collected to 2.9 Å resolution. The crystal belonged to space group P6122 or P6522, with unit-cell parameters a = b = 73.59, c = 157.66 Å, α = β = 90, γ = 120°.

scholarly journals Crystallization and preliminary crystallographic analysis of LipC12, a true lipase isolated through a metagenomics approach

2012 ◽  
Vol 68 (2) ◽  
pp. 175-177 ◽  
Author(s):  
V. P. Martini ◽  
A. Glogauer ◽  
J. Iulek ◽  
E. M. Souza ◽  
F. O. Pedrosa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Three Dimensional ◽  
Sodium Formate ◽  
Crystallographic Analysis ◽  
Dimensional Structure ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Sequence Identity

LipC12, a true lipase from family I.1 of bacterial lipases which was previously isolated through a metagenomics approach, contains 293 amino acids. Among lipases of known three-dimensional structure, it has a sequence identity of 47% to the lipase fromPseudomonas aeruginosaPAO1. Recombinant N-terminally His6-tagged LipC12 protein was expressed inEscherichia coli, purified in a homogenous form and crystallized in several conditions, with the best crystals being obtained using 2.0 Msodium formate and 0.1 Mbis-tris propane pH 7.0. X-ray diffraction data were collected to 2.70 Å resolution. The crystals belonged to the tetragonal space groupP4122, with unit-cell parametersa=b= 58.62,c = 192.60 Å.

scholarly journals Crystallization of SHARPIN using an automated two-dimensional grid screen for optimization

2012 ◽  
Vol 68 (7) ◽  
pp. 816-819 ◽  
Author(s):  
Benjamin Stieglitz ◽  
Katrin Rittinger ◽  
Lesley F. Haire
Keyword(s):  
Escherichia Coli ◽  
Protein Concentration ◽  
Unit Cell ◽  
Data Sets ◽  
Two Dimensional ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
One Step

An N-terminal fragment of human SHARPIN was recombinantly expressed inEscherichia coli, purified and crystallized. Crystals suitable for X-ray diffraction were obtained by a one-step optimization of seed dilution and protein concentration using a two-dimensional grid screen. The crystals belonged to the primitive tetragonal space groupP43212, with unit-cell parametersa = b = 61.55,c= 222.81 Å. Complete data sets were collected from native and selenomethionine-substituted protein crystals at 100 K to 2.6 and 2.0 Å resolution, respectively.

Purification, solution properties and crystallization of SIV integrase containing a continuous core and C-terminal domain

1999 ◽  
Vol 55 (11) ◽  
pp. 1906-1910 ◽  
Author(s):  
Ying Li ◽  
Youwei Yan ◽  
Joan Zugay-Murphy ◽  
Bei Xu ◽  
James L. Cole ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Solution Properties ◽  
Amino Acid Residues ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
Data Set ◽  
X Ray ◽  
Cell Parameters ◽  
Molecular Weights ◽  
Immunodeficiency Virus

The C-terminal two-thirds segment of integrase derived from the simian immunodeficiency virus has been cloned, expressed in Escherichia coli, and purified to greater than 95% homogeneity. The protein encompasses amino-acid residues 50–293 and contains a F185H substitution to enhance solubility. In dilute solutions at concentrations below 1 mg ml−1, the enzyme is predominantly dimeric. At the higher concentrations (>10 mg ml−1) required to enable crystallization, the enzyme self-associates to form species with molecular weights greater than 200 kDa. Despite the apparent high aggregation in solution, the enzyme crystallizes from a 8%(v/v) polyethylene glycol (molecular weight 6000) solution in a form suitable for X-ray diffraction studies. The resulting single crystals belong to the space group P212121, with unit-cell parameters a = 79.76, b = 99.98, c = 150.2 Å, α = β = γ = 90° and Z = 4. Under X-ray irradiation generated with a rotating-anode generator, the crystals diffract to 2.8 Å resolution and allow collection of a native 3 Å resolution diffraction data set.

scholarly journals Crystallization and preliminary X-ray diffraction analysis of orotate phosphoribosyltransferase from the human malaria parasitePlasmodium falciparum

2012 ◽  
Vol 68 (2) ◽  
pp. 244-246 ◽  
Author(s):  
Yasuhide Takashima ◽  
Eiichi Mizohata ◽  
Keiji Tokuoka ◽  
Sudaratana R. Krungkrai ◽  
Yukiko Kusakari ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Orotic Acid ◽  
Diffusion Method ◽  
Tetragonal Symmetry ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
Orotate Phosphoribosyltransferase ◽  
X Ray ◽  
Cell Parameters ◽  
Vapour Diffusion

Orotate phosphoribosyltransferase (OPRT) catalyzes the Mg2+-dependent condensation of orotic acid (OA) with 5-α-D-phosphorylribose 1-diphosphate (PRPP) to yield diphosphate (PPi) and the nucleotide orotidine 5′-monophosphate. OPRT fromPlasmodium falciparumproduced inEscherichia coliwas crystallized by the sitting-drop vapour-diffusion method in complex with OA and PRPP in the presence of Mg2+. The crystal exhibited tetragonal symmetry, belonging to space groupP41orP43, with unit-cell parametersa=b= 49.15,c = 226.94 Å. X-ray diffraction data were collected to 2.5 Å resolution at 100 K using a synchrotron-radiation source.

scholarly journals Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Zucchini fromDrosophila melanogaster

2012 ◽  
Vol 68 (11) ◽  
pp. 1346-1350
Author(s):  
Satoshi Fukuhara ◽  
Hiroshi Nishimasu ◽  
Luc Bonnefond ◽  
Naoki Matsumoto ◽  
Ryuichiro Ishitani ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Phospholipase D ◽  
Germ Cells ◽  
Underlying Mechanism ◽  
Crystallographic Analysis ◽  
Genomic Integrity ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters

PIWI-interacting RNAs (piRNAs) bind PIWI proteins and silence transposons to maintain the genomic integrity of germ cells. Zucchini (Zuc), a phospholipase D superfamily member, is conserved among animals and is implicated in piRNA biogenesis. However, the underlying mechanism by which Zuc participates in piRNA biogenesis remains elusive.Drosophila melanogasterZuc (DmZuc) was expressed inEscherichia coli, purified and crystallized. X-ray diffraction data were collected to 1.75 Å resolution. The crystal belonged to space groupP21, with unit-cell parametersa= 55.0,b= 71.2,c= 56.3 Å, β = 107.9°.

Structural Study of a new Compound Based on Acid 1,4-Cyclohexanedicarboxylic (1,4-CDC)

2010 ◽  
Vol 6 (1) ◽  
pp. 891-896
Author(s):  
Manel Halouani ◽  
M. Dammak ◽  
N. Audebrand ◽  
L. Ktari
Keyword(s):  
Aqueous Solution ◽  
Water Molecules ◽  
Carboxyl Group ◽  
X Ray Diffraction ◽  
Compound I ◽  
Unit Cell Parameters ◽  
Monoclinic System ◽  
X Ray ◽  
Cell Parameters ◽  
Cyclohexanedicarboxylic Acid

One nickel 1,4-cyclohexanedicarboxylate coordination polymers, Ni2 [(O10C6H4)(COO)2].2H2O  (I), was hydrothermally synthesized from an aqueous solution of Ni (NO3)2.6H2O, (1,4-CDC) (1,4-CDC = 1,4-cyclohexanedicarboxylic acid) and tetramethylammonium nitrate. Compound (I) crystallizes in the monoclinic system with the C2/m space group. The unit cell parameters are a = 20.1160 (16) Å, b = 9.9387 (10) Å, c = 6.3672 (6) Å, β = 97.007 (3) (°), V= 1263.5 (2) (Å3) and Dx= 1.751g/cm3. The refinement converged into R= 0.036 and RW = 0.092. The structure, determined by single crystal X-ray diffraction, consists of two nickel atoms Ni (1) and Ni (2). Lots of ways of which is surrounded by six oxygen atoms, a carboxyl group and two water molecules.

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