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Literature summary for 1.18.1.2 extracted from

  • Decottignies, P.; Flesch, V.; Gerard-Hirne, C.; Le Marechal, P.
    Role of positively charged residues in Chlamydomonas reinhardtii ferredoxin-NADP+-reductase (2003), Plant Physiol. Biochem., 41, 637-642.
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli, no posttranslational modification of the recombinant enzyme, the recombinant enzyme shows 26% of native enzyme activity Chlamydomonas reinhardtii

Protein Variants

Protein Variants Comment Organism
K135L site-directed mutagenesis, altered tertiary structure compared to the wild-type enzyme, no methylation at K83, 10% of native wild-type enzyme activity Chlamydomonas reinhardtii
K83L site-directed mutagenesis, altered tertiary structure compared to the wild-type enzyme, no methylation at K83, 2-4% of native wild-type enzyme activity Chlamydomonas reinhardtii
K83L/K89L site-directed mutagenesis, altered tertiary structure compared to the wild-type enzyme, no methylation at K83, 0.3% of native wild-type enzyme activity Chlamydomonas reinhardtii
K89L site-directed mutagenesis, altered tertiary structure compared to the wild-type enzyme, no methylation at K83, 4% of native wild-type enzyme activity Chlamydomonas reinhardtii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics Chlamydomonas reinhardtii
0.0026
-
reduced ferredoxin native enzyme, pH 8.0, 30°C Chlamydomonas reinhardtii
0.0066
-
reduced ferredoxin recombinant enzyme, pH 8.0, 30°C Chlamydomonas reinhardtii
0.007
-
NADPH native enzyme, pH 8.0, 30°C Chlamydomonas reinhardtii
0.033
-
NADPH recombinant enzyme, pH 8.0, 30°C Chlamydomonas reinhardtii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36500
-
native enzyme, gel filtration Chlamydomonas reinhardtii
37500
-
native enzyme, native PAGE Chlamydomonas reinhardtii
38500
-
recombinant enzyme, gel filtration Chlamydomonas reinhardtii
42000
-
recombinant enzyme, native PAGE Chlamydomonas reinhardtii

Organism

Organism UniProt Comment Textmining
Chlamydomonas reinhardtii
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
side-chain modification the native enzyme is posttranslationally trimethylated at lysine residues K83, K89, and K135 inducing changes in the tertiary enzyme structure Chlamydomonas reinhardtii

Reaction

Reaction Comment Organism Reaction ID
2 reduced ferredoxin + NADP+ + H+ = 2 oxidized ferredoxin + NADPH positively charged residues K83 and K89 are required for catalytic activity, methylation of K83, K89, and K135 is required for optimal enzyme activity Chlamydomonas reinhardtii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Chlamydomonas reinhardtii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,6-dichlorophenolindophenol + NADPH + H+
-
Chlamydomonas reinhardtii reduced 2,6-dichlorophenolindophenol + NADP+
-
r
NADPH + H+ + oxidized 2,6-dichlorophenolindophenol diaphorase activity Chlamydomonas reinhardtii NADP+ + reduced 2,6-dichlorophenolindophenol
-
?
reduced ferredoxin + NADP+ ferredoxin-dependent cytochrome c reduction, ferredoxin binding is independent of enzyme methylation status, positively charged residues K83 and K89 are required for activity Chlamydomonas reinhardtii oxidized ferredoxin + NADPH
-
?
reduced ferredoxin + NADP+
-
Chlamydomonas reinhardtii oxidized ferredoxin + NADPH + H+
-
r

Subunits

Subunits Comment Organism
monomer
-
Chlamydomonas reinhardtii

Synonyms

Synonyms Comment Organism
ferredoxin-NADP+-reductase
-
Chlamydomonas reinhardtii
ferredoxin-NADP-reductase
-
Chlamydomonas reinhardtii
FNR
-
Chlamydomonas reinhardtii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Chlamydomonas reinhardtii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
35
-
2,6-dichlorophenolindophenol native enzyme, pH 8.0, 30°C Chlamydomonas reinhardtii
37
-
2,6-dichlorophenolindophenol recombinant enzyme, pH 8.0, 30°C Chlamydomonas reinhardtii
90
-
reduced ferredoxin native enzyme, pH 8.0, 30°C Chlamydomonas reinhardtii
174
-
reduced ferredoxin recombinant enzyme, pH 8.0, 30°C Chlamydomonas reinhardtii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Chlamydomonas reinhardtii

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Chlamydomonas reinhardtii
NADPH
-
Chlamydomonas reinhardtii

pI Value

Organism Comment pI Value Maximum pI Value
Chlamydomonas reinhardtii native and recombinant enzyme, isoelectric focusing
-
5.8