Insulin crystallization: The route from hanging-drop vapour diffusion to controlled crystallization in droplet microfluidics

2022 ◽  
pp. 126516
Author(s):  
Joana Ferreira ◽  
Zsuzsa Sárkány ◽  
Filipa Castro ◽  
Fernando Rocha ◽  
Simon Kuhn
Keyword(s):  
Droplet Microfluidics ◽  
Hanging Drop ◽  
Vapour Diffusion ◽  
Controlled Crystallization

scholarly journals Crystallization and preliminary X-ray diffraction analysis of the Csu pili CsuC–CsuA/B chaperone–major subunit pre-assembly complex fromAcinetobacter baumannii

2015 ◽  
Vol 71 (6) ◽  
pp. 770-774 ◽  
Author(s):  
Natalia Pakharukova ◽  
Minna Tuittila ◽  
Sari Paavilainen ◽  
Anton Zavialov
Keyword(s):  
Diffusion Method ◽  
X Ray Diffraction ◽  
Hanging Drop ◽  
Unit Cell Parameters ◽  
Gram Negative ◽  
X Ray ◽  
Cell Parameters ◽  
Abiotic Surfaces ◽  
Vapour Diffusion ◽  
Fimbrial Adhesins

The attachment of many Gram-negative pathogens to biotic and abiotic surfaces is mediated by fimbrial adhesins, which are assembledviathe classical, alternative and archaic chaperone–usher (CU) pathways. The archaic CU fimbrial adhesins have the widest phylogenetic distribution, yet very little is known about their structure and mechanism of assembly. To elucidate the biogenesis of archaic CU systems, structural analysis of the Csu fimbriae, which are used byAcinetobacter baumanniito form stable biofilms and cause nosocomial infection, was focused on. The major fimbriae subunit CsuA/B complexed with the CsuC chaperone was purified from the periplasm ofEscherichia colicells co-expressing CsuA/B and CsuC, and the complex was crystallized in PEG 3350 solution using the hanging-drop vapour-diffusion method. Selenomethionine-labelled CsuC–CsuA/B complex was purified and crystallized under the same conditions. The crystals diffracted to 2.40 Å resolution and belonged to the hexagonal space groupP6422, with unit-cell parametersa=b= 94.71,c = 187.05 Å, α = β = 90, γ = 120°. Initial phases were derived from a single anomalous diffraction (SAD) experiment using the selenomethionine derivative.

scholarly journals Crystallization and preliminary X-ray analysis of formate oxidase, an enzyme of the glucose–methanol–choline oxidoreductase family

2010 ◽  
Vol 66 (9) ◽  
pp. 1064-1066 ◽  
Author(s):  
Yoshifumi Maeda ◽  
Daiju Doubayashi ◽  
Takumi Ootake ◽  
Masaya Oki ◽  
Bunzo Mikami ◽  
...  
Keyword(s):  
Carbon Dioxide ◽  
Hydrogen Peroxide ◽  
Single Crystal ◽  
Aspergillus Oryzae ◽  
Diffraction Data ◽  
Diffusion Method ◽  
Hanging Drop ◽  
Space Group ◽  
X Ray ◽  
Vapour Diffusion

Formate oxidase (FOD), which catalyzes the oxidation of formate to yield carbon dioxide and hydrogen peroxide, belongs to the glucose–methanol–choline oxidoreductase (GMCO) family. FOD fromAspergillus oryzaeRIB40, which has a modified FAD as a cofactor, was crystallized at 293 K by the hanging-drop vapour-diffusion method. The crystal was orthorhombic and belonged to space groupC2221. Diffraction data were collected from a single crystal to 2.4 Å resolution.

Vapour-diffusion protein crystallization in newly designed pore strips

2004 ◽  
Vol 37 (6) ◽  
pp. 862-866 ◽  
Author(s):  
Carien Dekker ◽  
Lesley Haire ◽  
Guy Dodson
Keyword(s):  
Phase Diagram ◽  
Nucleation Rate ◽  
Protein Crystallization ◽  
Hanging Drop ◽  
Crystal Forms ◽  
Kinetics Of Crystallization ◽  
Vapour Diffusion ◽  
Kinetics Of

Use of hanging drops in a pore rather than on cover slips increases the total surface accessible for evaporation. This will have an effect on the kinetics of crystallization and on the degree of convection, both of which could affect the nucleation rate. The crystallization of a set of well known proteins in pore strips has been tested. The results show that crystallization in pore strips is possible, and that nucleation is comparable with that in hanging drops despite the use of smaller volumes. Nonetheless, in about 25% of the cases, faster nucleation was observed. In a few cases, different crystal forms appeared, suggesting that pore crystallization may allow a different trajectory through the phase diagram compared with the hanging drop.

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 Bacillus subtilis regulatory protein GerE

1998 ◽  
Vol 54 (6) ◽  
pp. 1453-1455 ◽  
Author(s):  
Valérie M.-A. Ducros ◽  
James A. Brannigan ◽  
Richard J. Lewis ◽  
Anthony J. Wilkinson
Keyword(s):  
Gene Expression ◽  
Regulatory Protein ◽  
Diffusion Method ◽  
Monoclinic Space Group ◽  
Hanging Drop ◽  
Gene Encoding ◽  
Regulate Gene Expression ◽  
X Ray ◽  
Vapour Diffusion ◽  
Regulate Gene

GerE is the latest-acting of a series of factors which regulate gene expression in the mother cell during sporulation in Bacillus. The gene encoding GerE has been cloned from B. subtilis and overexpressed in Escherichia coli. Purified GerE has been crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The small plate-like crystals belong to the monoclinic space group C2 and diffract beyond 2.2 Å resolution with a synchrotron radiation X-ray source.

Crystallization and preliminary crystal analysis of yeast hexokinase PI and PII

1999 ◽  
Vol 55 (12) ◽  
pp. 2047-2048 ◽  
Author(s):  
Paula R. Kuser ◽  
Alexander M. Golubev ◽  
Igor Polikarpov
Keyword(s):  
Signal Transduction ◽  
Unit Cell ◽  
Hanging Drop ◽  
Synchrotron Diffraction ◽  
Unit Cell Parameters ◽  
Space Group ◽  
Cell Parameters ◽  
Vapour Diffusion ◽  
Hanging Drop Method ◽  
Yeast Hexokinase

Hexokinase is the prime enzyme of the Embden–Meyerhof pathway and is responsible for the first stage of energy conversion. It catalyzes the transfer of a phosphate to glucose to form glucose-6-phosphate. Yeast hexokinase PII is also known to play an important role in glucose signal transduction. Crystals of yeast hexokinase isoforms PI and PII were obtained by vapour-diffusion techniques using the hanging-drop method. Isoform PI crystals belong to the space group P212121, with unit-cell parameters a = 62.12, b = 78.87, c = 144.74 Å. Unit-cell parameters for isoform PII crystals are a = b = 142.81, c = 58.46 Å and the space group is I4. Synchrotron diffraction data have been collected to 2.2 Å resolution from the isoform PII crystal, whereas isoform PI diffracted to 3.1 Å.

scholarly journals Crystallization and crystallographic analysis of branching enzymes fromCyanothecesp. ATCC 51142

2015 ◽  
Vol 71 (8) ◽  
pp. 1109-1113 ◽  
Author(s):  
Mari Hayashi ◽  
Ryuichiro Suzuki ◽  
Christophe Colleoni ◽  
Steven G. Ball ◽  
Naoko Fujita ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Asymmetric Unit ◽  
Hanging Drop ◽  
Unit Cell Parameters ◽  
Cell Parameters ◽  
Cyanobacterial Species ◽  
Vapour Diffusion ◽  
Branching Enzymes

Several cyanobacterial species, includingCyanothecesp. ATCC 51142, remarkably have four isoforms of α-glucan branching enzymes (BEs). Based on their primary structures, they are classified into glycoside hydrolase (GH) family 13 (BE1, BE2 and BE3) or family 57 (GH57 BE). In the present study, GH13-type BEs fromCyanothecesp. ATCC 51142 (BE1, BE2 and BE3) have been overexpressed inEscherichia coliand biochemically characterized. The recombinant BE1 was crystallized by the hanging-drop vapour-diffusion method. Crystals of BE1 were obtained at 293 K in the presence of 0.2 MMg2+, 7–10%(w/v) ethanol, 0.1 MHEPES–NaOH pH 7.2–7.9. The crystals belonged to the tetragonal space groupP41212, with unit-cell parametersa = b = 133.75,c= 185.90 Å, and diffracted to beyond 1.85 Å resolution. Matthews coefficient calculations suggested that the crystals of BE1 contained two molecules in the asymmetric unit.

scholarly journals Crystallization and X-ray analysis ofD-threonine aldolase fromChlamydomonas reinhardtii

2017 ◽  
Vol 73 (2) ◽  
pp. 86-89
Author(s):  
Yuki Hirato ◽  
Masaru Goto ◽  
Mayumi Tokuhisa ◽  
Minoru Tanigawa ◽  
Katsushi Nishimura
Keyword(s):  
Diffusion Method ◽  
Asymmetric Unit ◽  
Hanging Drop ◽  
Unit Cell Parameters ◽  
Solvent Content ◽  
X Ray ◽  
Cell Parameters ◽  
Threonine Aldolase ◽  
Vapour Diffusion ◽  
Resolution Analysis

D-Threonine aldolase from the green algaChlamydomonas reinhardtii(CrDTA) catalyzes the interconversion of several β-hydroxy-D-amino acids (e.g.D-threonine) and glycine plus the corresponding aldehydes. Recombinant CrDTA was overexpressed inEscherichia coliand purified to homogeneity; it was subsequently crystallized using the hanging-drop vapour-diffusion method at 295 K. Data were collected and processed at 1.85 Å resolution. Analysis of the diffraction pattern showed that the crystal belonged to space groupP1, with unit-cell parametersa= 64.79,b= 74.10,c= 89.94 Å, α = 77.07, β = 69.34, γ = 71.93°. The asymmetric unit contained four molecules of CrDTA. The Matthews coefficient was calculated to be 2.12 Å3 Da−1and the solvent content was 41.9%.

Preliminary X-ray crystallographic studies of a newly defined human theta-class glutathione transferase

1998 ◽  
Vol 54 (1) ◽  
pp. 148-150 ◽  
Author(s):  
William J. McKinstry ◽  
Aaron J. Oakley ◽  
Jamie Rossjohn ◽  
Denis Verger ◽  
Kian-Leong Tan ◽  
...  
Keyword(s):  
Glutathione Transferase ◽  
Critical Role ◽  
Environmental Pollutants ◽  
Diffusion Method ◽  
Asymmetric Unit ◽  
Hanging Drop ◽  
X Ray ◽  
Cell Dimensions ◽  
Vapour Diffusion ◽  
Glutathione S Transferases

Human theta-class glutathione S-transferases (GST's) appear to play a critical role in the metabolism of a variety of environmental pollutants but in some cases the products of the reaction are carcinogenic. Crystals of a human theta-class GST, namely hGSTT2-2, have been grown from polyethylene glycol by the hanging-drop vapour-diffusion method. The crystals belong to the trigonal space group P3121 with cell dimensions of a = b = 94.0 and c = 120.5 Å. They contain two monomers in the asymmetric unit and diffract to 3.0 Å resolution.

scholarly journals Plant-specific DUF1110 protein fromOryza sativa: expression, purification and crystallization

2016 ◽  
Vol 72 (6) ◽  
pp. 480-484 ◽  
Author(s):  
Kenichi Harada ◽  
Eiki Yamashita ◽  
Kento Inoue ◽  
Koji Yamaguchi ◽  
Toshimichi Fujiwara ◽  
...  
Keyword(s):  
Diffusion Method ◽  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Hanging Drop ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Sequence Identity ◽  
Vapour Diffusion ◽  
Domain Of Unknown Function

The Os01T0156300 protein fromOryza sativahas been classified into the domain of unknown function (DUF) family DUF1110. DUF1110 family members exist in monocotyledons but not in dicotyledons, and share no sequence identity with proteins for which structures have been reported. In this study, the Os01T0156300 protein was crystallized using the hanging-drop vapour-diffusion method. X-ray diffraction data were collected to 1.84 Å resolution. The crystal belonged to space groupP21, with unit-cell parametersa= 89.9,b= 89.8,c= 107.1 Å, β = 106.6°. The asymmetric unit was estimated to contain 6–11 molecules.

Sign in / Sign up

Export Citation Format

Share Document