Preliminary X-ray crystallographic analysis of a plant defence protein, the polygalacturonase-inhibiting protein from Phaseolus vulgaris

2000 ◽  
Vol 56 (1) ◽  
pp. 98-100 ◽  
Author(s):  
Andrew Leech ◽  
Benedetta Mattei ◽  
Luca Federici ◽  
Giuli De Lorenzo ◽  
Andrew M. Hemmings
Keyword(s):  
Amino Acid ◽  
Phaseolus Vulgaris ◽  
Plant Defence ◽  
Crystallographic Analysis ◽  
Leucine Rich Repeat ◽  
Asymmetric Unit ◽  
X Ray ◽  
Vapour Diffusion ◽  
Polygalacturonase Inhibiting Protein ◽  
Inhibiting Protein

A leucine-rich repeat plant protein involved in resistance to pathogens, a polygalacturonase-inhibiting protein (PGIP-1) from Phaseolus vulgaris, has been crystallized and preliminary X-ray characterization has been performed. The protein contains ten repeats of a short (24 amino-acid) leucine-rich repeat motif. Single crystals of the protein were grown from vapour-diffusion experiments using PEG 2K monomethylether as precipitant; these crystals diffract to at least 2.3 Å resolution. The space group is P21, with two molecules of PGIP-1 in the asymmetric unit; the crystals contain approximately 38% solvent.

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.

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%.

scholarly journals Expression, purification, crystallization and preliminary X-ray analysis of tannase fromLactobacillus plantarum

2013 ◽  
Vol 69 (4) ◽  
pp. 456-459 ◽  
Author(s):  
Mingbo Wu ◽  
Xiaohong Peng ◽  
Hua Wen ◽  
Qin Wang ◽  
Qianming Chen ◽  
...  
Keyword(s):  
Synchrotron Radiation ◽  
Gallic Acid ◽  
Diffusion Method ◽  
Ester Bond ◽  
Asymmetric Unit ◽  
Data Set ◽  
X Ray ◽  
Vapour Diffusion ◽  
Serine Esterases ◽  
Hydrolysis Of

Tannase catalyses the hydrolysis of the galloyl ester bond of tannins to release gallic acid. It belongs to the serine esterases and has wide applications in the food, feed, beverage, pharmaceutical and chemical industries. The tannase fromLactobacillus plantarumwas cloned, expressed and purified. The protein was crystallized by the sitting-drop vapour-diffusion method with microseeding. The crystals belonged to space groupP1, with unit-cell paramtersa= 46.5,b= 62.8,c= 83.8 Å, α = 70.4, β = 86.0, γ = 79.4°. Although the enzyme exists mainly as a monomer in solution, it forms a dimer in the asymmetric unit of the crystal. The crystals diffracted to beyond 1.60 Å resolution using synchrotron radiation and a complete data set was collected to 1.65 Å resolution.

Polygalacturonase-inhibiting protein (PGIP) in plant defence: a structural view

2006 ◽  
Vol 67 (6) ◽  
pp. 528-533 ◽  
Author(s):  
Adele Di Matteo ◽  
Daniele Bonivento ◽  
Demetrius Tsernoglou ◽  
Luca Federici ◽  
Felice Cervone
Keyword(s):  
Plant Defence ◽  
Polygalacturonase Inhibiting Protein ◽  
Inhibiting Protein

scholarly journals Crystallization and preliminary X-ray crystallographic analysis of human myotubularin-related protein 1

2015 ◽  
Vol 71 (3) ◽  
pp. 261-265 ◽  
Author(s):  
Seoung Min Bong ◽  
Seung Won Yang ◽  
Ji-Woong Choi ◽  
Seung Jun Kim ◽  
Byung Il Lee
Keyword(s):  
Escherichia Coli ◽  
Polyethylene Glycol ◽  
Synchrotron Radiation ◽  
Crystallographic Analysis ◽  
Asymmetric Unit ◽  
Related Protein ◽  
Unit Cell Parameters ◽  
Solvent Content ◽  
X Ray ◽  
Cell Parameters

Myotubularin-related protein 1 is a phosphatase that dephosphorylates phospholipids such as phosphatidylinositol 3-phosphate or phosphatidylinositol 3,5-bisphosphate. In this study, human MTMR1 was overexpressed inEscherichia coli, purified and crystallized at 277 K using polyethylene glycol 20 000 as a precipitant. Diffraction data were collected to 2.0 Å resolution using synchrotron radiation. The crystals belonged to space groupP1, with unit-cell parametersa= 67.219,b= 96.587,c= 97.581 Å, α = 87.597, β = 86.072, γ = 77.327°. Assuming the presence of four molecules in the asymmetric unit, the calculated Matthews coefficient value was 2.61 Å3 Da−1and the corresponding solvent content was 52.9%.

scholarly journals Characterization, crystallization and preliminary X-ray crystallographic analysis of the Uba5 fragment necessary for high-efficiency activation of Ufm1

2014 ◽  
Vol 70 (6) ◽  
pp. 765-768 ◽  
Author(s):  
Shutao Xie
Keyword(s):  
High Efficiency ◽  
Crystallographic Analysis ◽  
Terminal Region ◽  
Adenylation Domain ◽  
Asymmetric Unit ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Multicellular Organisms

Uba5 is the smallest ubiquitin-like molecule-activating enzyme and contains an adenylation domain and a C-terminal region. This enzyme only exists in multicellular organisms. The mechanism through which the enzyme recognizes and activates ubiquitin-fold modifier 1 (Ufm1) remains unknown. In this study, Uba5 adenylation domains with different C-terminal region lengths were cloned, expressed and purified. The results of anin vitrotruncation assay suggest that Uba5 residues 57–363 comprise the minimal fragment required for the high-efficiency activation of Ufm1. Crystallization of Uba5 residues 57–363 was performed at 277 K using PEG 3350 as the precipitant, and crystals optimized by microseeding diffracted to 2.95 Å resolution, with unit-cell parametersa=b= 97.66,c= 144.83 Å, α = β = 90, γ = 120°. There is one molecule in the asymmetric unit; the Matthews coefficient and the solvent content were calculated to be 2.93 Å3 Da−1and 58.1%, respectively.

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