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Literature summary extracted from

  • Georgescauld, F.; Moynie, L.; Habersetzer, J.; Dautant, A.
    Structure of Mycobacterium tuberculosis nucleoside diphosphate kinase R80N mutant in complex with citrate (2014), Acta Crystallogr. Sect. F, 70, 40-43.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.4.6 purified recombinant enzyme mutant R80N, mixing of 200 nl of 11 mg/ml protein in 20 mM Tris-HCl, pH 7.5, and 20 mM MgCl2 with 200 nl of reservoir solution containing 30% w/v PEG 4000, 0.2 M ammonium acetate, and 0.1 M trisodium citrate, pH 5.6, a few hours, 20°C, X-ray diffraction structure determination and analysis at 1.9 A resolution, molecular replacement method using the wild-type Mt-NDPK structure as search model, PDB ID 1k44, modelling Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
2.7.4.6 D93N site-directed mutagenesis, the mutation breaks the intersubunit salt bridge Arg80-Asp93 which contributes to the thermal stability of the hexamer. The mutant thermal stability dramatically decreases by 27.6°C to 48.4°C, compared to 76°C for the wild-type enzyme Mycobacterium tuberculosis
2.7.4.6 R80N site-directed mutagenesis, the mutation affects the intersubunit salt bridge Arg80-Asp93 which contributes to the thermal stability of the hexamer. The mutant thermal stability dramatically decreases by 8.0°C to 68°C, compared to 76°C for the wild-type enzyme. In mutant R80N, the salt bridge is replaced by intersubunit hydrogen bonds that contribute to the thermal stability of the hexamer Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
2.7.4.6 Mycobacterium tuberculosis
-
-
-

Subunits

EC Number Subunits Comment Organism
2.7.4.6 hexamer
-
 Mycobacterium tuberculosis
2.7.4.6 More the intersubunit salt bridge Arg80-Asp93 contributes to the thermal stability of the hexamer Mycobacterium tuberculosis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.4.6 76
-
 wild-type enzyme, stable up to, the intersubunit salt bridge Arg80-Asp93 contributes to the thermal stability of the hexamer Mycobacterium tuberculosis