Literature summary for 1.18.1.5 extracted from

Sevrioukova, I.F.; Poulos, T.L.
Arginines 65 and 310 in putidaredoxin reductase are critical for interaction with putidaredoxin (2010), Biochemistry, 49, 5160-5166.

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structure of a covalently linked putidaredoxin reductase (Pdr)-putidaredoxin (Pdx) complex. Residues R65 and R310 are the key elements required for the formation of a productive electron transfer complex with Pdx. The C-terminal lysine cluster assists in Pdx docking by fine-tuning Pdr-Pdx interactions to achieve the optimal geometry between the redox centers, and the basic surface residues in Pdr-like ferredoxin reductases not only define specificity for the redox partner but also may facilitate its dissociation	Pseudomonas putida

Crystallization (Comment)

Organism

crystal structure of a covalently linked putidaredoxin reductase (Pdr)-putidaredoxin (Pdx) complex. Residues R65 and R310 are the key elements required for the formation of a productive electron transfer complex with Pdx. The C-terminal lysine cluster assists in Pdx docking by fine-tuning Pdr-Pdx interactions to achieve the optimal geometry between the redox centers, and the basic surface residues in Pdr-like ferredoxin reductases not only define specificity for the redox partner but also may facilitate its dissociation

Pseudomonas putida

Protein Variants

Protein Variants	Comment	Organism
K339A	moderate decrease in the binding affinity and reduction of Pdx	Pseudomonas putida
K387A	moderate decrease in the binding affinity and reduction of Pdx	Pseudomonas putida
K409A	moderate decrease in the binding affinity and reduction of Pdx	Pseudomonas putida
N384A	mutation has no effect on the Pdr-Pdx interaction	Pseudomonas putida
R310A	mutation lowers the interprotein electron tranfer rate by 20-30fold without perturbing the Pdx association step	Pseudomonas putida
R310E	mutation decreases both the Pdr-to-Pdx ET and partner binding affinity by 100- and 8fold, respectively	Pseudomonas putida
R65A	mutation lowers the interprotein electron tranfer rate by 20-30fold without perturbing the Pdx association step	Pseudomonas putida

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.017	-	NADH	mutant R65A, pH 7.5, 25°C, cosubstrate dichlorophenolindophenol	Pseudomonas putida
0.018	-	NADH	mutant R310A, pH 7.5, 25°C, cosubstrate dichlorophenolindophenol	Pseudomonas putida
0.022	-	NADH	mutant R339A, pH 7.5, 25°C, cosubstrate dichlorophenolindophenol	Pseudomonas putida
0.025	-	NADH	mutant R339A, pH 7.5, 25°C, cosubstrate K3Fe(CN)6	Pseudomonas putida
0.027	-	NADH	wild-type, pH 7.5, 25°C, cosubstrate dichlorophenolindophenol	Pseudomonas putida
0.029	-	NADH	mutant K409A, pH 7.5, 25°C, cosubstrate K3Fe(CN)6	Pseudomonas putida
0.029	-	oxidized 2,6-dichlorophenolindolphenol	mutant R65A, pH 7.5, 25°C	Pseudomonas putida
0.031	-	NADH	mutant K409A, pH 7.5, 25°C, cosubstrate dichlorophenolindophenol	Pseudomonas putida
0.034	-	NADH	mutant R65A, pH 7.5, 25°C, cosubstrate K3Fe(CN)6	Pseudomonas putida
0.036	-	NADH	mutant K387A, pH 7.5, 25°C, cosubstrate K3Fe(CN)6	Pseudomonas putida
0.036	-	Fe(CN)63-	mutant K409A, pH 7.5, 25°C	Pseudomonas putida
0.036	-	NADH	mutant R310A, pH 7.5, 25°C, cosubstrate K3Fe(CN)6	Pseudomonas putida
0.037	-	NADH	mutant R310E, pH 7.5, 25°C, cosubstrate dichlorophenolindophenol	Pseudomonas putida
0.037	-	NADH	mutant R310E, pH 7.5, 25°C, cosubstrate K3Fe(CN)6	Pseudomonas putida
0.038	-	NADH	wild-type, pH 7.5, 25°C, cosubstrate K3Fe(CN)6	Pseudomonas putida
0.04	-	Fe(CN)63-	wild-type, pH 7.5, 25°C	Pseudomonas putida
0.04	-	Fe(CN)63-	mutant R310E, pH 7.5, 25°C	Pseudomonas putida
0.042	-	NADH	mutant K387A, pH 7.5, 25°C, cosubstrate dichlorophenolindophenol	Pseudomonas putida
0.043	-	Fe(CN)63-	mutant N384A, pH 7.5, 25°C	Pseudomonas putida
0.044	-	Fe(CN)63-	mutant K387A, pH 7.5, 25°C	Pseudomonas putida
0.045	-	Fe(CN)63-	mutant R339A, pH 7.5, 25°C	Pseudomonas putida
0.05	-	NADH	mutant N384A, pH 7.5, 25°C, cosubstrate dichlorophenolindophenol	Pseudomonas putida
0.05	-	NADH	mutant N384A, pH 7.5, 25°C, cosubstrate K3Fe(CN)6	Pseudomonas putida
0.05	-	Fe(CN)63-	mutant R65A, pH 7.5, 25°C	Pseudomonas putida
0.083	-	oxidized 2,6-dichlorophenolindolphenol	mutant K387A, pH 7.5, 25°C	Pseudomonas putida
0.083	-	oxidized 2,6-dichlorophenolindolphenol	mutant N384A, pH 7.5, 25°C	Pseudomonas putida
0.091	-	oxidized 2,6-dichlorophenolindolphenol	mutant K409A, pH 7.5, 25°C	Pseudomonas putida
0.097	-	oxidized 2,6-dichlorophenolindolphenol	wild-type, pH 7.5, 25°C	Pseudomonas putida
0.125	-	oxidized 2,6-dichlorophenolindolphenol	mutant R310A, pH 7.5, 25°C	Pseudomonas putida
0.125	-	oxidized 2,6-dichlorophenolindolphenol	mutant R339A, pH 7.5, 25°C	Pseudomonas putida
0.143	-	Fe(CN)63-	mutant R310A, pH 7.5, 25°C	Pseudomonas putida
0.2	-	oxidized 2,6-dichlorophenolindolphenol	mutant R310E, pH 7.5, 25°C	Pseudomonas putida

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	P16640	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2 ferricytochrome c + NADH	-	Pseudomonas putida	2 ferrocytochrome c + NAD+ + H+	-	?
NADH + H+ + Fe(CN)63-	-	Pseudomonas putida	NAD+ + Fe(CN)62-	-	?
NADH + H+ + oxidized 2,6-dichlorophenolindophenol	-	Pseudomonas putida	NAD+ + reduced 2,6-dichlorophenolindophenol	-	?
reduced putidaredoxin + NAD+	-	Pseudomonas putida	oxidized putidaredoxin + NADH + H+	-	?

Synonyms

Synonyms	Comment	Organism
Pdr	-	Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
203	-	oxidized 2,6-dichlorophenolindolphenol	mutant R65A, pH 7.5, 25°C	Pseudomonas putida
283	-	oxidized 2,6-dichlorophenolindolphenol	mutant R310E, pH 7.5, 25°C	Pseudomonas putida
288	-	oxidized 2,6-dichlorophenolindolphenol	mutant R339A, pH 7.5, 25°C	Pseudomonas putida
358	-	oxidized 2,6-dichlorophenolindolphenol	mutant N384A, pH 7.5, 25°C	Pseudomonas putida
377	-	oxidized 2,6-dichlorophenolindolphenol	mutant K409A, pH 7.5, 25°C	Pseudomonas putida
385	-	oxidized 2,6-dichlorophenolindolphenol	wild-type, pH 7.5, 25°C	Pseudomonas putida
408	-	oxidized 2,6-dichlorophenolindolphenol	mutant K387A, pH 7.5, 25°C	Pseudomonas putida
543	-	Fe(CN)63-	mutant R339A, pH 7.5, 25°C	Pseudomonas putida
560	-	Fe(CN)63-	mutant R65A, pH 7.5, 25°C	Pseudomonas putida
617	-	oxidized 2,6-dichlorophenolindolphenol	mutant R310A, pH 7.5, 25°C	Pseudomonas putida
650	-	NADH	mutant R339A, pH 7.5, 4°C	Pseudomonas putida
720	-	NADH	mutant R310E, pH 7.5, 4°C	Pseudomonas putida
729	-	NADH	mutant R310A, pH 7.5, 4°C	Pseudomonas putida
754	-	NADH	mutant R65A, pH 7.5, 4°C	Pseudomonas putida
755	-	Fe(CN)63-	mutant K409A, pH 7.5, 25°C	Pseudomonas putida
772	-	Fe(CN)63-	mutant K387A, pH 7.5, 25°C	Pseudomonas putida
780	-	NADH	mutant K387A, pH 7.5, 4°C	Pseudomonas putida
790	-	Fe(CN)63-	mutant R310E, pH 7.5, 25°C	Pseudomonas putida
800	-	NADH	mutant K409A, pH 7.5, 4°C	Pseudomonas putida
806	-	NADH	wild-type, pH 7.5, 4°C	Pseudomonas putida
820	-	NADH	mutant N384A, pH 7.5, 4°C	Pseudomonas putida
898	-	Fe(CN)63-	mutant N384A, pH 7.5, 25°C	Pseudomonas putida
982	-	Fe(CN)63-	wild-type, pH 7.5, 25°C	Pseudomonas putida
1485	-	Fe(CN)63-	mutant R310A, pH 7.5, 25°C	Pseudomonas putida