Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.85 extracted from

  • Qu, C.; Akanuma, S.; Tanaka, N.; Moriyama, H.; Oshima, T.
    Design, X-ray crystallography, molecular modelling and thermal stability studies of mutant enzymes at site 172 of 3-isopropylmalate dehydrogenase from Thermus thermophilus (2001), Acta Crystallogr. Sect. D, 57, 225-232.
    View publication on PubMed

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzymes A172V, A172G and A172F. As in the case of A172L enzyme, the A172F mutant can not be crystallized by the salting-out technique with ammonium sulfate. The crystal is obtained at pH 4.8 using polyethylene glycol 4000 as a precipitant. Crystals of mutant enzyme A172E are obtained from a drop equilibrated with reservoir solution consisting of 0.8 M ammonium sulfate pH 6.0 at either 15°C or 20°C K. Two types of crystals: one hexagonal bipyramidal and the other is tetragonal Thermus thermophilus

Protein Variants

Protein Variants Comment Organism
A172D melting temperature is reduced by 2.7°C compared to wild-type enzyme Thermus thermophilus
A172F mutation causes rearrangement in domain structure, which leads to a higher thermostability compared to wild-type enzyme Thermus thermophilus
A172F melting temperature is increased by 1.8°C compared to wild-type enzyme Thermus thermophilus
A172G melting temperature is reduced by 0.3°C compared to wild-type enzyme, crystal structure is similar to that of the wild-type enzyme Thermus thermophilus
A172I melting temperature is increased by 2°C compared to wild-type enzyme Thermus thermophilus
A172L mutation causes rearrangement in domain structure, which leads to a higher thermostability compared to wild-type enzyme Thermus thermophilus
A172L melting temperature is increased by 3°C compared to wild-type enzyme Thermus thermophilus
A172V melting temperature is increased by 1°C compared to wild-type enzyme, crystal structure is similar to that of the wild-type enzyme Thermus thermophilus

Organism

Organism UniProt Comment Textmining
Thermus thermophilus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2R,3S)-3-isopropylmalate + NAD+
-
Thermus thermophilus 2-oxoisocaproate + NADH + H+ + CO2
-
?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
-
Thermus thermophilus
61
-
midpoint of thermal unfolding curve, mutant enzyme A172D Thermus thermophilus
64
-
midpoint of thermal unfolding curve, mutant enzyme A172G Thermus thermophilus
65
-
midpoint of thermal unfolding curve, mutant enzyme A172V Thermus thermophilus
66
-
midpoint of thermal unfolding curve, mutant enzyme A172I and A172E Thermus thermophilus
67
-
midpoint of thermal unfolding curve, mutant enzyme A172L and A172F Thermus thermophilus
87
-
clear correlation was observed between the hyrophobicity and the thermostability of the enzyme Thermus thermophilus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Thermus thermophilus