Literature summary for 2.7.4.6 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.

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Protein Variants	Comment	Organism
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
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

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	-	-	-

Subunits

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

Temperature Stability [°C]

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