DNA ligase (NAD+) / DNA ligase (NAD+) activity / base-excision repair, DNA ligation / DNA ligation / NAD+ binding / DNA replication / DNA binding / metal ion binding / cytosol Similarity search - Function
Laminin - #30 / Laminin / DNA ligase-like, N-terminal NAD+-binding domain / Dna Ligase; domain 1 - #70 / Zinc-finger, NAD-dependent DNA ligase C4-type / NAD-dependent DNA ligase C4 zinc finger domain / NAD-dependent DNA ligase, active site / NAD-dependent DNA ligase, conserved site / NAD-dependent DNA ligase signature 1. / NAD-dependent DNA ligase signature 2. ...Laminin - #30 / Laminin / DNA ligase-like, N-terminal NAD+-binding domain / Dna Ligase; domain 1 - #70 / Zinc-finger, NAD-dependent DNA ligase C4-type / NAD-dependent DNA ligase C4 zinc finger domain / NAD-dependent DNA ligase, active site / NAD-dependent DNA ligase, conserved site / NAD-dependent DNA ligase signature 1. / NAD-dependent DNA ligase signature 2. / NAD-dependent DNA ligase / NAD-dependent DNA ligase, OB-fold / NAD-dependent DNA ligase, adenylation / NAD-dependent DNA ligase, N-terminal / NAD-dependent DNA ligase adenylation domain / NAD-dependent DNA ligase OB-fold domain / Ligase N family / DisA/LigA, helix-hairpin-helix motif / Helix-hairpin-helix motif / DNA ligase/mRNA capping enzyme / Helix hairpin bin / RuvA domain 2-like / Other non-globular / Helix-hairpin-helix domain / BRCA1 C Terminus (BRCT) domain / D-amino Acid Aminotransferase; Chain A, domain 1 / breast cancer carboxy-terminal domain / Helix-hairpin-helix DNA-binding motif, class 1 / Helix-hairpin-helix DNA-binding motif class 1 / BRCT domain profile. / BRCT domain / BRCT domain superfamily / Nucleic acid-binding proteins / Dna Ligase; domain 1 / 5' to 3' exonuclease, C-terminal subdomain / Special / DNA polymerase; domain 1 / OB fold (Dihydrolipoamide Acetyltransferase, E2P) / Helix Hairpins / Nucleic acid-binding, OB-fold / Beta Barrel / 2-Layer Sandwich / Orthogonal Bundle / Mainly Beta / Mainly Alpha / Alpha Beta Similarity search - Domain/homology
Mass: 18.015 Da / Num. of mol.: 320 / Source method: isolated from a natural source / Formula: H2O
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Experimental details
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Experiment
Experiment
Method: X-RAY DIFFRACTION / Number of used crystals: 1
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Sample preparation
Crystal
Density Matthews: 2.52 Å3/Da / Density % sol: 51.28 %
Crystal grow
Temperature: 295 K / Method: vapor diffusion, sitting drop / pH: 6 Details: EcoLigA (0.3 mM) was reacted with 5 mM MgCl2 and 0.3 mM NAD+ for 30 min at 22 C. The ligase-adenylylation reaction was then quenched by adding 10 mM EDTA. The mixture was supplemented with ...Details: EcoLigA (0.3 mM) was reacted with 5 mM MgCl2 and 0.3 mM NAD+ for 30 min at 22 C. The ligase-adenylylation reaction was then quenched by adding 10 mM EDTA. The mixture was supplemented with 26-bp nicked duplex DNA (0.318 mM). This LigA-nucleic acid solution was mixed 1:2 with a well solution containing 200 mM ammonium sulfate, 50 mM sodium acetate, 24% PEG-4000. Crystals were grown at 22 C by the sitting drop vapor diffusion method. Crystals appeared after 3 days. The crystals were transferred serially to solutions containing 5% glycerol/24% PEG-4000, 10% glycerol/26% PEG-4000, 15% glycerol/28% PEG-4000, and 20% glycerol/30% PEG-4000 in 100 mM ammonium sulfate, 50 mM sodium acetate, after which they were flash-frozen in liquid nitrogen. , pH 6.0, VAPOR DIFFUSION, SITTING DROP, temperature 295K
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