scholarly journals Purification, crystallization and preliminary crystallographic analysis of a ribosome-recycling factor fromThermoanaerobacter tengcongensis(TteRRF)

2014 ◽  
Vol 70 (5) ◽  
pp. 588-591 ◽  
Author(s):  
Guijun Shang ◽  
Duo Feng ◽  
Fang Lu ◽  
Hongjie Zhang ◽  
Huaixing Cang ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Elongation Factor ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Ribosome Recycling Factor ◽  
Reductive Methylation ◽  
Data Set ◽  
Cell Parameters ◽  
Thermoanaerobacter Tengcongensis ◽  
Fourth Step

Ribosome-recycling factor (RRF) plays an essential role in the fourth step of protein synthesis in prokaryotes. RRF combined with elongation factor G (EF-G) disassembles the post-termination ribosome complex and recycles the protein synthesis machine for the next round of translation. A reductive-methylation-modified RRF fromThermoanaerobacter tengcongensis(TteRRF) has been crystallized using the vapour-diffusion method. The crystal grew in a condition consisting of 0.1 Mcitric acid pH 3.5, 3.0 MNaCl and 50 mg ml−1methylated protein solution at 289 K. A complete data set was collected from a crystal to 2.80 Å resolution using synchrotron radiation at 100 K. The crystal belonged to space groupP6122/P6522 with unit-cell parametersa=b= 103.26,c= 89.17 Å. The asymmetric unit was estimated to contain one molecule ofTteRRF.

scholarly journals Cloning, purification and preliminary crystallographic analysis of theHelicobacter pyloripseudaminic acid biosynthesisN-acetyltransferase PseH

2014 ◽  
Vol 70 (9) ◽  
pp. 1276-1279 ◽  
Author(s):  
Yu C. Liu ◽  
Abu I. Ud-Din ◽  
Anna Roujeinikova
Keyword(s):  
Crystallographic Analysis ◽  
Diffusion Method ◽  
X Ray Diffraction ◽  
Data Set ◽  
Cell Parameters ◽  
Nonulosonic Acid ◽  
Pseudaminic Acid ◽  
The Common ◽  
H Pylori ◽  
Development Of Gastric Cancer

Helicobacter pyloriinfection is the common cause of gastritis and duodenal and stomach ulcers, which have been linked to a higher risk of the development of gastric cancer. The motility that facilitates persistent infection requires functional flagella that are heavily glycosylated with 5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (pseudaminic acid). Pseudaminic acid biosynthesis protein H (PseH) catalyzes the third step in its biosynthetic pathway, producing UDP-2,4-diacetamido-2,4,6-trideoxy-β-L-altropyranose. Crystals ofH. pyloriPseH have been grown by the hanging-drop vapour-diffusion method using diammonium tartrate as a precipitating agent. The crystals belonged to space groupI222 orI212121, with unit-cell parametersa= 107.8,b= 145.4,c= 166.3 Å. A complete X-ray diffraction data set has been collected to 2.5 Å resolution using cryocooling conditions and synchrotron radiation.

scholarly journals Crystallization and preliminary X-ray crystallographic analysis of Pz peptidase B fromGeobacillus collagenovoransMO-1

2012 ◽  
Vol 68 (7) ◽  
pp. 757-759 ◽  
Author(s):  
Hiroaki Nakano ◽  
Allin Hosokawa ◽  
Ryuji Tagawa ◽  
Koji Inaka ◽  
Kazunori Ohta ◽  
...  
Keyword(s):  
Space Station ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Unit Cell Parameters ◽  
Data Set ◽  
Peptide Substrates ◽  
X Ray ◽  
Cell Parameters ◽  
Experiment Module ◽  
Counter Diffusion

Pz peptidase B is an intracellular M3 metallopeptidase that is found together with Pz peptidase A in the thermophileGeobacillus collagenovoransMO-1 and recognizes collagen-specific tripeptide units (-Gly-Pro-X-). These peptidases have low homology in their primary structures; however, their cleavage patterns towards peptide substrates are similar. In this work, Pz peptidase B was crystallized using the counter-diffusion method. Data were collected to a resolution of 1.6 Å at 100 K from a crystal obtained in the Japanese Experiment Module (JEM; also known as `Kibo') at the International Space Station (ISS). The crystal belonged to the trigonal space groupP3121, with unit-cell parametersa=b= 87.64,c= 210.5 Å. A complete data set was also obtained from crystals of selenomethionine-substituted protein.

scholarly journals Expression, purification and preliminary crystallographic analysis of the cryptic polo-box domain ofCaenorhabditis elegansZYG-1

2014 ◽  
Vol 70 (10) ◽  
pp. 1346-1350
Author(s):  
Ekaterina Shimanovskaya ◽  
Gang Dong
Keyword(s):  
Crystallographic Analysis ◽  
Structural Knowledge ◽  
Diffusion Method ◽  
Lysine Methylation ◽  
Wild Type ◽  
Data Set ◽  
Cell Parameters ◽  
Anomalous Data ◽  
Strain Bl21 ◽  
Shed Light

ZYG-1 is a polo-like kinase essential for centriole assembly inCaenorhabditis elegans. The targeting of ZYG-1 to nascent centrioles isviaits central cryptic polo-box (CPB) domain. To shed light on the molecular basis of ZYG-1 recruitment, it is necessary to obtain structural knowledge of the ZYG-1 CPB. Here, the expression, purification and preliminary crystallographic analysis of the ZYG-1 CPB are reported. The protein was overexpressed inEscherichia colistrain BL21 (DE3), purified by multi-step chromatography and crystallized using the vapour-diffusion method. Crystals of the wild-type protein exhibited an order–disorder pathology, which was solved by reductive lysine methylation. A complete anomalous data set was collected to 2.54 Å resolution at the Se Kedge (λ = 0.9792 Å). The crystal belonged to space groupP2, with unit-cell parametersa= 53.3,b= 60.09,c= 87.51 Å, β = 93.31°. There were two molecules in the asymmetric unit.

scholarly journals Cloning, expression, refolding, purification and preliminary crystallographic analysis of the sensory domain of theCampylobacterchemoreceptor for aspartate A (CcaA)

2015 ◽  
Vol 71 (1) ◽  
pp. 110-113 ◽  
Author(s):  
Mayra A. Machuca ◽  
Yu C. Liu ◽  
Anna Roujeinikova
Keyword(s):  
Campylobacter Jejuni ◽  
Inclusion Bodies ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Hanging Drop ◽  
Unit Cell Parameters ◽  
Data Set ◽  
Cell Parameters ◽  
Extracellular Signals ◽  
Denatured Protein

InCampylobacter jejuni, chemotaxis and motility have been identified as important virulence factors that are required for host colonization and invasion. Chemotactic recognition of extracellular signals is mediated by the periplasmic sensory domains of its transducer-like proteins (Tlps). In this study, the sensory domain of theC. jejunichemoreceptor for aspartate A (CcaA) has been expressed inEscherichia coliand purified from inclusion bodies. The urea-denatured protein was refolded and then crystallized by the hanging-drop vapour-diffusion method using PEG 3350 as a precipitating agent. A complete data set has been collected to 1.4 Å resolution using cryocooling conditions and synchrotron radiation. The crystals belonged to space groupP1, with unit-cell parametersa= 39.3,b= 43.3,c= 50.9 Å, α = 92.5, β = 111.4, γ = 114.7°.

scholarly journals Recombinant ACHT1 fromArabidopsis thaliana: crystallization and X-ray crystallographic analysis

2017 ◽  
Vol 73 (7) ◽  
pp. 382-385 ◽  
Author(s):  
Weimin Pan ◽  
Junchao Wang ◽  
Ye Yang ◽  
Lin Liu ◽  
Min Zhang
Keyword(s):  
Escherichia Coli ◽  
Arabidopsis Thaliana ◽  
Disulfide Bonds ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Unit Cell Parameters ◽  
Data Set ◽  
X Ray ◽  
Cell Parameters ◽  
Vapour Diffusion

Thioredoxins (Trxs) play important roles in chloroplasts by linking photosynthetic light reactions to a series of plastid functions. They execute their function by regulating the oxidation and reduction of disulfide bonds. ACHT1 (atypical cysteine/histidine-rich Trx1) is a thylakoid-associated thioredoxin-type protein found in theArabidopsis thalianachloroplast. Recombinant ACHT1 protein was overexpressed inEscherichia coli, purified and crystallized by the vapour-diffusion method. The crystal diffracted to 1.7 Å resolution and a complete X-ray data set was collected. Preliminary crystallographic analysis suggested that the crystals belonged to space groupC2221, with unit-cell parametersa = 102.7,b= 100.6,c= 92.8 Å.

scholarly journals Cloning, refolding, purification and preliminary crystallographic analysis of the sensory domain of theCampylobacterchemoreceptor for multiple ligands (CcmL)

2015 ◽  
Vol 71 (2) ◽  
pp. 211-216 ◽  
Author(s):  
Mayra A. Machuca ◽  
Yu C. Liu ◽  
Simone A. Beckham ◽  
Anna Roujeinikova
Keyword(s):  
Polyethylene Glycol ◽  
Campylobacter Jejuni ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Hanging Drop ◽  
Unit Cell Parameters ◽  
Data Set ◽  
Cell Parameters ◽  
Vapour Diffusion ◽  
Multiple Ligands

A periplasmic sensory domain of theCampylobacter jejunichemoreceptor for multiple ligands (CcmL) has been crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 3350 as a precipitating agent. A complete data set was collected to 1.3 Å resolution using cryocooling conditions and synchrotron radiation. The crystals belonged to space groupP21, with unit-cell parametersa= 42.6,b= 138.0,c= 49.0 Å, β = 94.3°.

scholarly journals Preparation, purification, crystallization and preliminary crystallographic analysis of dual-domain β-propeller phytase fromBacillussp. HJB17

2014 ◽  
Vol 70 (12) ◽  
pp. 1671-1674 ◽  
Author(s):  
Fang Lu ◽  
Gangxin Guo ◽  
Qianqian Li ◽  
Duo Feng ◽  
Yong Liu ◽  
...  
Keyword(s):  
Fold Increase ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Sodium Acetate Trihydrate ◽  
Data Set ◽  
Cell Parameters ◽  
Acetate Trihydrate ◽  
C Terminus ◽  
Phytate Hydrolysis ◽  
Dual Domain

β-Propeller phytases (BPPs) are abundant in nature. Recently, dual-domain BPPs have been found in which the typical BPP domain is responsible for phytate hydrolysis. The dual-domain BPP (PhyH) fromBacillussp. HJB17 was obtained with an incomplete N-terminal BPP domain (PhyH-DI; residues 41–318) and a typical BPP domain (PhyH-DII; residues 319–644) at the C-terminus. PhyH-DI was found to act synergistically (with a 1.2–2.5-fold increase in phosphate release) with PhyH-DII, other BPPs (PhyP and 168PhyA) and a histidine acid phosphatase. The structure of PhyH was therefore studied with the aim of explaining these functions. PhyH with the secreted signal peptide of the first 40 amino acids deleted (PhyHT) was cloned and expressed inEscherichia coli. Purified and active PhyHT protein was obtained by refolding from the precipitant. PhyHT was crystallized using the vapour-diffusion method. The crystal grew in a condition consisting of 0.2 Msodium acetate trihydrate, 0.1 MTris–HCl pH 9.5, 25%(w/v) polyethylene glycol 4000 using 1 mg ml−1protein solution at 289 K. A complete data set was collected from a crystal to 2.85 Å resolution using synchrotron radiation at 100 K. The crystal belonged to space groupP1211, with unit-cell parametersa= 46.82,b= 140.19,c= 81.94 Å, α = 90.00, β = 92.00, γ = 90.00°. The asymmetric unit was estimated to contain one molecule of PhyHT.

scholarly journals Crystallographic analysis of FAD-dependent glucose dehydrogenase

2015 ◽  
Vol 71 (8) ◽  
pp. 1017-1019 ◽  
Author(s):  
Hirofumi Komori ◽  
Koji Inaka ◽  
Naoki Furubayashi ◽  
Michinari Honda ◽  
Yoshiki Higuchi
Keyword(s):  
Aspergillus Terreus ◽  
Glucose Dehydrogenase ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Asymmetric Unit ◽  
Unit Cell Parameters ◽  
Data Set ◽  
Solvent Content ◽  
X Ray ◽  
Cell Parameters

An FAD-dependent glucose dehydrogenase (GDH) fromAspergillus terreuswas purified and crystallized at 293 K using the sitting-drop vapour-diffusion method. A data set was collected to a resolution of 1.6 Å from a single crystal at 100 K using a rotating-anode X-ray source. The crystal belonged to space groupP21, with unit-cell parametersa= 56.56,b= 135.74,c= 74.13 Å, β = 90.37°. The asymmetric unit contained two molecules of GDH. The Matthews coefficient was calculated to be 2.2 Å3 Da−1and the solvent content was estimated to be 44%.

Crystallization and preliminary X-ray analysis of Thermotoga maritima ribosome recycling factor

1999 ◽  
Vol 55 (12) ◽  
pp. 2049-2050 ◽  
Author(s):  
Maria Selmer ◽  
Salam Al-Karadaghi ◽  
Go Hirokawa ◽  
Akira Kaji ◽  
Anders Liljas
Keyword(s):  
Protein Synthesis ◽  
Diffraction Data ◽  
Thermotoga Maritima ◽  
Asymmetric Unit ◽  
Ribosome Recycling Factor ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Fourth Step ◽  
Prokaryotic Protein

Thermotoga maritima ribosome recycling factor (RRF) is one of the proteins catalyzing the fourth step in prokaryotic protein synthesis, ribosome recycling. The RRF protein was crystallized with ammonium sulfate. Native diffraction data to 2.55 Å resolution were obtained at the MAX II synchrotron from a flash-frozen crystal at 100 K. The crystals belong to space group P41212 or P43212, with unit-cell parameters a = b = 47, c = 298 Å, and probably contain one monomer per asymmetric unit.

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