Crystallization and preliminary X-ray diffraction analysis of anL-amino-acid oxidase fromBothrops jararacussuvenom

Snake-venom L-amino-acid oxidases (SV-LAAOs) trigger a wide range of local and systematic effects, including inhibition of platelet aggregation, cytotoxicity, haemolysis, apoptosis and haemorrhage. These effects mainly arise from the uncontrolled release of the hydrogen peroxide that is produced by the redox reaction involving L-amino acids catalyzed by these flavoenzymes. Taking their clinical relevance into account, few SV-LAAOs have been structurally characterized and the structural determinants responsible for their broad direct and indirect pharmacological activities remain unclear. In this work, an LAAO fromBothrops jararacussuvenom (BJu-LAAO) was purified and crystallized. The BJu-LAAO crystals belonged to space groupP21, with unit-cell parametersa = 66.38,b= 72.19,c= 101.53 Å, β = 90.9°. The asymmetric unit contained two molecules and the structure was determined and partially refined at 3.0 Å resolution.

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Isolation, crystallization and preliminary X-ray diffraction analysis ofL-amino-acid oxidase fromVipera ammodytes ammodytesvenom

Acta Crystallographica Section F Structural Biology and Crystallization Communications ◽

10.1107/s1744309108027036 ◽

2008 ◽

Vol 64 (10) ◽

pp. 918-921 ◽

Cited By ~ 3

Author(s):

Dessislava Georgieva ◽

Anna Kardas ◽

Friedrich Buck ◽

Markus Perbandt ◽

Christian Betzel

Keyword(s):

Amino Acid ◽

Diffraction Analysis ◽

Acid Oxidase ◽

X Ray Diffraction ◽

Amino Acid Oxidase ◽

X Ray

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Crystallization and preliminary X-ray diffraction studies of anL-amino-acid oxidase fromLachesis mutavenom

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x14017877 ◽

2014 ◽

Vol 70 (11) ◽

pp. 1556-1559 ◽

Cited By ~ 4

Author(s):

Anwar Ullah ◽

Rehana Masood ◽

Patrick Jack Spencer ◽

Mário Tyago Murakami ◽

Raghuvir Krishnaswamy Arni

Keyword(s):

Amino Acid ◽

Rapid Evolution ◽

Model Systems ◽

Acid Oxidase ◽

X Ray Diffraction ◽

Amino Acid Oxidase ◽

Cell Parameters ◽

Structural Modifications ◽

Lachesis Muta ◽

Venom Proteins

Snake-venom proteins form multi-component defence systems by the recruitment and rapid evolution of nonvenomous proteins and hence serve as model systems to understand the structural modifications that result in toxicity. L-Amino-acid oxidases (LAAOs) are encountered in a number of snake venoms and have been implicated in the inhibition of platelet aggregation, cytotoxicity, haemolysis, apoptosis and haemorrhage. An L-amino-acid oxidase fromLachesis mutavenom has been purified and crystallized. The crystals belonged to space groupP21, with unit-cell parametersa= 66.05,b= 79.41,c= 100.52 Å, β = 96.55°. The asymmetric unit contained two molecules and the structure has been determined and partially refined at 3.0 Å resolution.

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An Automated Detection System for Most L-α-Amino Acids

Clinical Chemistry ◽

10.1093/clinchem/15.2.154 ◽

1969 ◽

Vol 15 (2) ◽

pp. 154-161 ◽

Cited By ~ 6

Author(s):

K Van Dyke ◽

C Szustkiewicz

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Detection System ◽

Automated System ◽

Acid Oxidase ◽

Enzymatic Reactions ◽

Amino Acid Oxidase ◽

Automated Method ◽

Wide Range

Abstract An automated system for the determination of the L-α form of the majority of amino acids is presented. The method is based upon oxidative deamination of the amino acid coupled with oxidation of o-dianisidine by hydrogen peroxide. This procedure can be used comparatively for the determination of a mixture of L-α-amino acids or for the majority of separated L-α-amino acids (especially in conjunction with column separations from urine and blood which give falsely positive identification with ninhydrin detection). The stereospecific nature of the L-α-amino acid oxidase enables the investigator to quantitate the amount of L-α-amino acid in the presence of the D-α form. From an academic viewpoint, the extreme sensitivity and wide range of the detection system make it advantageous for the study of the enzyme itself. This automated method also may be employed to follow enzymatic reactions—e.g., those catalyzed by peptidases or racemases. The methodology is extremely convenient with good reagent stability and is much more sensitive than manometric technics.

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Crystallization and preliminary X-ray diffraction analysis of the homing endonuclease I-CvuI fromChlorella vulgarisin complex with its target DNA

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x1400065x ◽

2014 ◽

Vol 70 (2) ◽

pp. 256-259 ◽

Cited By ~ 2

Author(s):

Pilar Redondo ◽

Nekane Merino ◽

Maider Villate ◽

Francisco J. Blanco ◽

Guillermo Montoya ◽

...

Keyword(s):

Diffraction Analysis ◽

Homing Endonuclease ◽

Homing Endonucleases ◽

Resolution Limit ◽

X Ray Diffraction ◽

Orthorhombic Space Group ◽

Strand Breaks ◽

X Ray ◽

Cell Parameters ◽

Wide Range

Homing endonucleases are highly specific DNA-cleaving enzymes that recognize long stretches of DNA. The engineering of these enzymes provides novel instruments for genome modification in a wide range of fields, including gene targeting, by inducing specific double-strand breaks. I-CvuI is a homing endonuclease from the green algaChlorella vulgaris. This enzyme was purified after overexpression inEscherichia coli. Crystallization experiments of I-CvuI in complex with its DNA target in the presence of Mg2+yielded crystals suitable for X-ray diffraction analysis. The crystals belonged to the orthorhombic space groupP212121, with unit-cell parametersa= 62.83,b= 83.56,c= 94.40 Å. The self-rotation function and the Matthews coefficient suggested the presence of one protein–DNA complex per asymmetric unit. The crystals diffracted to a resolution limit of 1.9 Å using synchrotron radiation.

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Limited Proteolysis and X-ray Crystallography Reveal the Origin of Substrate Specificity and of the Rate-Limiting Product Release during Oxidation ofd-Amino Acids Catalyzed by Mammaliand-Amino Acid Oxidase†,‡

Biochemistry ◽

10.1021/bi963023s ◽

1997 ◽

Vol 36 (19) ◽

pp. 5624-5632 ◽

Cited By ~ 32

Author(s):

Maria A. Vanoni ◽

Antonio Cosma ◽

Daniela Mazzeo ◽

Andrea Mattevi ◽

Flavia Todone ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Substrate Specificity ◽

Limited Proteolysis ◽

Acid Oxidase ◽

Amino Acid Oxidase ◽

X Ray ◽

Product Release ◽

X Ray Crystallography ◽

Rate Limiting

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Crystallization and preliminary X-ray diffraction analysis ofBacillus subtilisYwfE, anL-amino-acid ligase

Acta Crystallographica Section F Structural Biology and Crystallization Communications ◽

10.1107/s174430911105425x ◽

2012 ◽

Vol 68 (2) ◽

pp. 203-206 ◽

Cited By ~ 2

Author(s):

Takeo Tsuda ◽

Tomomi Suzuki ◽

Shuichi Kojima

Keyword(s):

Amino Acid ◽

Diffusion Method ◽

Dependent Manner ◽

Asymmetric Unit ◽

X Ray Diffraction ◽

Hanging Drop ◽

Unit Cell Parameters ◽

Data Set ◽

X Ray ◽

Cell Parameters

Bacillus subtilisYwfE, an L-amino-acid ligase, catalyzes the formation of an α-dipeptide from L-amino acids in an ATP-dependent manner. In order to elucidate the substrate-recognition mode and the reaction mechanism of this ligase, native and selenomethionine-derivatized (SeMet) crystals of YwfE in the presence of ADP, MgCl2and the dipeptide L-Ala-L-Gln were obtained using the hanging-drop vapour-diffusion method. These crystals diffracted to 1.9 and 2.8 Å resolution, respectively. Preliminary SAD phase calculations using the data set from the SeMet crystal suggested that the crystal belonged to the hexagonal space groupP6522, with unit-cell parametersa=b= 90.85,c = 250.31 Å, and contained one molecule in the asymmetric unit with a solvent content of 57.3%.

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Purification, crystallization and preliminary X-ray diffraction analysis of theN-acetyltransferase SAV0826 fromStaphylococcus aureus

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x13034493 ◽

2014 ◽

Vol 70 (2) ◽

pp. 211-214

Author(s):

Parul Srivastava ◽

Yogesh B. Khandokar ◽

Jade K. Forwood

Keyword(s):

Diffraction Analysis ◽

Single Molecule ◽

Protein Function ◽

Bacterial Species ◽

Diffusion Method ◽

X Ray Diffraction ◽

X Ray ◽

Cell Parameters ◽

Wide Range ◽

Antibiotic Drug

Staphylococcus aureusis a prevalent microorganism that is capable of causing a wide range of infections and diseases. Several strains of this bacterial species have developed antibiotic resistance to methicillin and vancomycin, and higher death rates are still being reported each year owing to multidrug-resistant strains. Certain GCN5-relatedN-acetyltransferases (GNATs) exhibit a broad substrate range, including aminoglycosides, histones, other proteins and serotonin, and have been implicated in antibiotic drug resistance. Here, the expression, purification, crystallization and preliminary X-ray diffraction analysis of a GNAT fromS. aureus(SaNAT) are reported. SaNAT was recombinantly expressed and crystallized by the hanging-drop vapour-diffusion method at 296 K, and the crystals diffracted to 1.7 Å resolution on the MX2 beamline at the Australian Synchrotron. The crystals belonged to space groupP43212, with unit-cell parametersa=b= 84.86,c= 49.06 Å, α = β = γ = 90°. A single molecule is likely to be present in the asymmetric unit. A full structural and functional analysis is currently being undertaken to provide novel insights into the protein function, which in turn may provide a basis for drug design.

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X-ray powder diffraction analysis of a new palladium(II) amino acid complex

Powder Diffraction ◽

10.1154/1.1648317 ◽

2004 ◽

Vol 19 (3) ◽

pp. 270-271 ◽

Cited By ~ 2

Author(s):

Pedro P. Corbi ◽

Antonio C. Massabni ◽

Claudio M. Costa-Neto

Keyword(s):

Amino Acid ◽

Diffraction Analysis ◽

Powder Diffraction ◽

Diffraction Data ◽

X Ray Diffraction ◽

Acid Complex ◽

X Ray ◽

Cell Parameters ◽

Amino Acid Complex ◽

Orthorhombic Cell

Powder X-ray diffraction data for a new palladium(II) amino acid complex, of composition PdC12H20N2O4S2, are presented in this paper. Orthorhombic cell parameters are: a=10.740 Å, b=19.999 Å, and c=5.2470 Å. © 2004 International Centre for Diffraction Data.

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