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Literature summary for 1.1.1.88 extracted from

  • Friesen, J.A.; Lawrence, C.M.; Stauffacher, C.V.; Rodwell, V.W.
    Structural determinants of nucleotide coenzyme specificity in the distinctive dinucleotide binding fold of HMG-CoA reductase from Pseudomonas mevalonii (1996), Biochemistry, 35, 11945-11950.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 Pseudomonas mevalonii

Protein Variants

Protein Variants Comment Organism
D146A 6670-fold more specific for NADP+ than wild-type Pseudomonas mevalonii
D146A/L148K 72200-fold more specific for NADP+ than wild-type Pseudomonas mevalonii
D146A/L148R 83300-fold more specific for NADP+ than wild-type Pseudomonas mevalonii
D146A/Q147K no activity with NADP+ as cofactor Pseudomonas mevalonii
D146A/T192K no activity with NADP+ as cofactor Pseudomonas mevalonii
D146A/T192R no activity with NADP+ as cofactor Pseudomonas mevalonii
D146G 1170-fold more specific for NADP+ than wild-type Pseudomonas mevalonii
D146G/L148K 55600-fold more specific for NADP+ than wild-type Pseudomonas mevalonii
D146G/L148R no activity with NADP+ as cofactor Pseudomonas mevalonii
D146G/T192K 3170-fold more specific for NADP+ than wild-type Pseudomonas mevalonii
D146G/T192R 4500-fold more specific for NADP+ than wild-type Pseudomonas mevalonii
D146N no activity with NADP+ as cofactor Pseudomonas mevalonii
D146S no activity with NADP+ as cofactor Pseudomonas mevalonii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.21
-
NAD+ wild-type enzyme, oxidative acylation of mevalonate Pseudomonas mevalonii
52
-
NADP+ wild-type enzyme, oxidative acylation of mevalonate Pseudomonas mevalonii

Organism

Organism UniProt Comment Textmining
Pseudomonas mevalonii
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-mevalonate + CoA + NADP+
-
Pseudomonas mevalonii 3-hydroxy-3-methylglutaryl-CoA + NADPH + H+
-
r

Cofactor

Cofactor Comment Organism Structure
additional information very detailed kinetic studies of mutants concerning NAD+/NADP+-affinities, the best specificity improvement is achieved by the D146A/L148R-mutant Pseudomonas mevalonii
NAD+
-
Pseudomonas mevalonii
NADH reverse reaction Pseudomonas mevalonii
NADP+ NAD+ is 600000fold more efficient than NADP+ in wild-type enzyme Pseudomonas mevalonii