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Literature summary for 2.7.4.6 extracted from

  • Georgescauld, F.; Moynie, L.; Habersetzer, J.; Cervoni, L.; Mocan, I.; Borza, T.; Harris, P.; Dautant, A.; Lascu, I.
    Intersubunit ionic interactions stabilize the nucleoside diphosphate kinase of Mycobacterium tuberculosis (2013), PLoS ONE, 8, e57867.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.4.6

Cloned(Commentary)

Cloned (Comment) Organism
mutant enzyme D93N is expressed in Escherichia coli BL21-CodonPlus(DE3)-RIL cells Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzyme D93N crystallizes to hexagonal plates with 2.0 M ammonium sulfate, 0.1 M Tris-HCl, pH 8.5, and to rods with 2.0 M ammonium sulfate, 2% (w/v) PEG400, 0.1 M HEPES, pH 7.5 Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
D93N the mutation dramatically decreases protein thermal stability (Tm value of 43.4°C) but is still hexameric and retains full activity. The mutant hexamer is stable and active up to 2.5 M guanidine hydrochloride Mycobacterium tuberculosis

General Stability

General Stability Organism
guanidinium and many other salts stabilize the hexamer of mutant D93N with a half maximum effect of about 0.1 M, increasing protein thermostability Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
Urea inactivation occurs at very low urea concentrations (0.5 M) Mycobacterium tuberculosis

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WJH7
-
-
Mycobacterium tuberculosis H37Rv P9WJH7
-
-

Purification (Commentary)

Purification (Comment) Organism
Q-Sepharose column chromatography Mycobacterium tuberculosis

Renatured (Commentary)

Renatured (Comment) Organism
the wild type enzyme unfolds and inactivates simultaneously in the presence of 4 M guanidine hydrochloride as with 4 M urea as denaturant. Refolding occurs at much lower guanidine hydrochloride concentrations (0.5-1 M) Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 8-bromoinosine 5'-diphosphate
-
 Mycobacterium tuberculosis ADP + 8-bromoinosine 5'-triphosphate
-
 ?
ATP + 8-bromoinosine 5'-diphosphate
-
 Mycobacterium tuberculosis H37Rv ADP + 8-bromoinosine 5'-triphosphate
-
 ?

Subunits

Subunits Comment Organism
hexamer x-ray crystallography Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
NDPK
-
 Mycobacterium tuberculosis
nucleoside diphosphate kinase
-
 Mycobacterium tuberculosis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
76
-
 the wild type enzyme has a temperature of half denaturation of 76°C Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.159
-
 8-bromoinosine 5'-diphosphate wild type enzyme, at pH 7.5 and 22°C Mycobacterium tuberculosis
0.23
-
 8-bromoinosine 5'-diphosphate mutant enzyme D93N, at pH 7.5 and 22°C Mycobacterium tuberculosis
232
-
 ATP mutant enzyme D93N, at pH 7.5 and 22°C Mycobacterium tuberculosis
264
-
 ATP wild type enzyme, at pH 7.5 and 22°C Mycobacterium tuberculosis