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Literature summary for 1.6.2.2 extracted from

  • Kimura, S.; Nishida, H.; Iyanagi, T.
    Effects of flavin-binding motif amino acid mutations in the NADH-cytochrome b5 reductase catalytic domain on protein stability and catalysis (2001), J. Biochem., 130, 481-490.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type, K97A, K97R, S99A, S99T, S99V, R63A, R63Q, R63K, Y65A and Y65F mutant enzymes in Escherichia coli Sus scrofa

Protein Variants

Protein Variants Comment Organism
K97A mutation in flavin-binding motif Sus scrofa
K97R mutation in flavin-binding motif Sus scrofa
R63A mutation in flavin-binding motif Sus scrofa
R63K mutation in flavin-binding motif Sus scrofa
R63Q mutation in flavin-binding motif Sus scrofa
S99A mutation in flavin-binding motif Sus scrofa
S99T mutation in flavin-binding motif Sus scrofa
S99V mutation in flavin-binding motif Sus scrofa
Y65A mutation in flavin-binding motif Sus scrofa
Y65F mutation in flavin-binding motif Sus scrofa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00148
-
ferricytochrome b5 S99A mutant enzyme Sus scrofa
0.0019
-
NADH S99T mutant enzyme Sus scrofa
0.0021
-
NADH R63K mutant enzyme Sus scrofa
0.0029
-
NADH K97R mutant enzyme Sus scrofa
0.003
-
NADH Y65F mutant enzyme Sus scrofa
0.0031
-
NADH recombinant wild-type enzyme Sus scrofa
0.0042
-
NADH Y65A mutant enzyme Sus scrofa
0.0044
-
NADH K97A mutant enzyme Sus scrofa
0.005
-
ferricytochrome b5 Y65A mutant enzyme Sus scrofa
0.0062
-
ferricytochrome b5 Y65F mutant enzyme Sus scrofa
0.0069
-
ferricytochrome b5 K97R mutant enzyme Sus scrofa
0.0073
-
ferricytochrome b5 R63K mutant enzyme Sus scrofa
0.0094
-
NADH S99V mutant enzyme Sus scrofa
0.0099
-
ferricytochrome b5 S99V mutant enzyme Sus scrofa
0.0104
-
ferricytochrome b5 recombinant wild-type enzyme Sus scrofa
0.0118
-
ferricytochrome b5 S99T mutant enzyme Sus scrofa
0.0161
-
ferricytochrome b5 K97A mutant enzyme Sus scrofa
0.0185
-
ferricytochrome b5 R63Q mutant enzyme Sus scrofa
0.021
-
NADH R63Q mutant enzyme Sus scrofa
0.026
-
NADH S99A mutant enzyme Sus scrofa
0.0421
-
ferricytochrome b5 R63A mutant enzyme Sus scrofa
0.104
-
NADH R63A mutant enzyme Sus scrofa

Organism

Organism UniProt Comment Textmining
Sus scrofa
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type, K97A, K97R, S99A, S99T, S99V, R63A, R63Q, R63K, Y65A and Y65F mutant enzymes Sus scrofa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
NADH + ferricytochrome b5
-
Sus scrofa NAD+ + H+ + ferrocytochrome b5
-
r

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
105
-
ferricytochrome b5 R63Q mutant enzyme Sus scrofa
107
-
ferricytochrome b5 Y65A mutant enzyme Sus scrofa
121
-
ferricytochrome b5 R63A mutant enzyme Sus scrofa
165
-
ferricytochrome b5 S99V mutant enzyme Sus scrofa
180
-
NADH Y65A mutant enzyme Sus scrofa
191
-
ferricytochrome b5 K97A mutant enzyme Sus scrofa
295
-
ferricytochrome b5 S99A mutant enzyme Sus scrofa
305
-
ferricytochrome b5 Y65F mutant enzyme Sus scrofa
347
-
ferricytochrome b5 R63K mutant enzyme Sus scrofa
375
-
NADH S99V mutant enzyme Sus scrofa
403
-
NADH R63A mutant enzyme Sus scrofa
412
-
NADH R63Q mutant enzyme Sus scrofa
415
-
ferricytochrome b5 K97R mutant enzyme Sus scrofa
435
-
NADH S99A mutant enzyme Sus scrofa
501
-
NADH Y65F mutant enzyme Sus scrofa
515
-
NADH R63K mutant enzyme Sus scrofa
520
-
ferricytochrome b5 S99T mutant enzyme Sus scrofa
661
-
ferricytochrome b5 recombinant wild-type enzyme Sus scrofa
851
-
NADH S99T mutant enzyme Sus scrofa
909
-
NADH K97R mutant enzyme Sus scrofa
1060
-
NADH K97A mutant enzyme Sus scrofa
1100
-
NADH recombinant wild-type enzyme Sus scrofa