Literature summary for 3.1.3.11 extracted from

Feng, L.; Sun, Y.; Deng, H.; Li, D.; Wan, J.; Wang, X.; Wang, W.; Liao, X.; Ren, Y.; Hu, X.
Structural and biochemical characterization of fructose-1,6/sedoheptulose-1,7-bisphosphatase from the cyanobacterium Synechocystis strain 6803 (2014), FEBS J., 281, 916-926.

Crystallization (Commentary)

Crystallization (Comment)	Organism
bifunctional D-fructose 1,6-bisphosphatase/sedoheptulose 1,7-bisphosphatase in complex with AMP and fructose 1,6-bisphosphate, to 2.3 A resolution. The allosteric inhibitor AMP and the substrate fructose 1,6-bisphosphate exhibit an unusual binding mode when in complex with D-fructose 1,6-bisphosphatase/sedoheptulose 1,7-bisphosphatase. AMP binds to the allosteric sites near the interface across the up/down subunit pairs C1C4 and C2C3 in the center of the tetramer, while fructose 1,6-bisphophate binds opposite to the interface between the horizontal subunit pairs C1C2 or C3C4. Formation of a disulfide linkage may occur between Cys75 and Cys99. The enzyme may be regulated through ligand binding and alteration of the structure of the enzyme complex	Synechocystis sp.

Crystallization (Comment)

Organism

bifunctional D-fructose 1,6-bisphosphatase/sedoheptulose 1,7-bisphosphatase in complex with AMP and fructose 1,6-bisphosphate, to 2.3 A resolution. The allosteric inhibitor AMP and the substrate fructose 1,6-bisphosphate exhibit an unusual binding mode when in complex with D-fructose 1,6-bisphosphatase/sedoheptulose 1,7-bisphosphatase. AMP binds to the allosteric sites near the interface across the up/down subunit pairs C1C4 and C2C3 in the center of the tetramer, while fructose 1,6-bisphophate binds opposite to the interface between the horizontal subunit pairs C1C2 or C3C4. Formation of a disulfide linkage may occur between Cys75 and Cys99. The enzyme may be regulated through ligand binding and alteration of the structure of the enzyme complex

Synechocystis sp.

Protein Variants

Protein Variants	Comment	Organism
D198H	more than 10fold reduction in kcat value, reduction in IC50 value of AMP	Synechocystis sp.
D200A	simultaneous decrease in substrate affinity and kcat value	Synechocystis sp.
D97A	almost complete loss of activity	Synechocystis sp.
E100A	almost complete loss of activity	Synechocystis sp.
E225A	almost complete loss of activity	Synechocystis sp.
E57A	almost complete loss of activity	Synechocystis sp.
F309A	greatly reduced activity	Synechocystis sp.
H215A	greatly reduced activity	Synechocystis sp.
K134A	greatly reduced activity, reduction in IC50 value of AMP	Synechocystis sp.
K29A	2fold increase in activity	Synechocystis sp.
N213A	greatly reduced activity	Synechocystis sp.
R164A	greatly reduced activity	Synechocystis sp.
R176A	simultaneous decrease in substrate affinity and kcat value	Synechocystis sp.
R178G	greatly reduced activity	Synechocystis sp.
R307A	greatly reduced activity	Synechocystis sp.
R314A	greatly reduced activity	Synechocystis sp.
T102A	more than 10fold reduction in kcat value, reduction in IC50 value of AMP	Synechocystis sp.
Y131A	greatly reduced activity	Synechocystis sp.

Inhibitors

Inhibitors	Comment	Organism	Structure
AMP	-	Synechocystis sp.

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.08	-	D-fructose 1,6-bisphosphate	mutant N213A, Hill coefficient 1.8, pH 8.0, 28°C	Synechocystis sp.
0.08	-	D-fructose 1,6-bisphosphate	mutant R314A, Hill coefficient 1.8, pH 8.0, 28°C	Synechocystis sp.
0.08	-	D-fructose 1,6-bisphosphate	wild-type, Hill coefficient 1.9, pH 8.0, 28°C	Synechocystis sp.
0.1	-	D-fructose 1,6-bisphosphate	mutant H215A, Hill coefficient 1.4, pH 8.0, 28°C	Synechocystis sp.
0.1	-	D-fructose 1,6-bisphosphate	mutant T102A, Hill coefficient 2.5, pH 8.0, 28°C	Synechocystis sp.
0.11	-	D-fructose 1,6-bisphosphate	mutant R164A, Hill coefficient 2.1, pH 8.0, 28°C	Synechocystis sp.
0.15	-	D-fructose 1,6-bisphosphate	mutant K29A, Hill coefficient 2.0, pH 8.0, 28°C	Synechocystis sp.
0.17	-	D-fructose 1,6-bisphosphate	mutant D198H, Hill coefficient 1.9, pH 8.0, 28°C	Synechocystis sp.
0.21	-	D-fructose 1,6-bisphosphate	mutant F309A, Hill coefficient 1.8, pH 8.0, 28°C	Synechocystis sp.
0.24	-	D-fructose 1,6-bisphosphate	mutant R307A, Hill coefficient 1.6, pH 8.0, 28°C	Synechocystis sp.
0.26	-	D-fructose 1,6-bisphosphate	mutant Y131A, Hill coefficient 1.6, pH 8.0, 28°C	Synechocystis sp.
0.5	-	D-fructose 1,6-bisphosphate	mutant K134A, Hill coefficient 2.2, pH 8.0, 28°C	Synechocystis sp.
0.61	-	D-fructose 1,6-bisphosphate	mutant R178A, Hill coefficient 1.1, pH 8.0, 28°C	Synechocystis sp.
1.32	-	D-fructose 1,6-bisphosphate	mutant D200A, Hill coefficient 1.4, pH 8.0, 28°C	Synechocystis sp.
3.2	-	D-fructose 1,6-bisphosphate	mutant R176A, Hill coefficient 1.2, pH 8.0, 28°C	Synechocystis sp.

Organism

Organism	UniProt	Comment	Textmining
Synechocystis sp.	P73922	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-fructose 1,6-bisphosphate + H2O	-	Synechocystis sp.	D-fructose 6-phosphate + phosphate	-	?

Synonyms

Synonyms	Comment	Organism
slr2094	-	Synechocystis sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.63	-	D-fructose 1,6-bisphosphate	mutant D200A, Hill coefficient 1.4, pH 8.0, 28°C	Synechocystis sp.
0.7	-	D-fructose 1,6-bisphosphate	mutant R176A, Hill coefficient 1.2, pH 8.0, 28°C	Synechocystis sp.
0.7	-	D-fructose 1,6-bisphosphate	mutant T102A, Hill coefficient 2.5, pH 8.0, 28°C	Synechocystis sp.
0.84	-	D-fructose 1,6-bisphosphate	mutant D198H, Hill coefficient 1.9, pH 8.0, 28°C	Synechocystis sp.
1.2	-	D-fructose 1,6-bisphosphate	mutant H215A, Hill coefficient 1.4, pH 8.0, 28°C	Synechocystis sp.
1.2	-	D-fructose 1,6-bisphosphate	mutant Y131A, Hill coefficient 1.6, pH 8.0, 28°C	Synechocystis sp.
1.8	-	D-fructose 1,6-bisphosphate	mutant K134A, Hill coefficient 2.2, pH 8.0, 28°C	Synechocystis sp.
2.3	-	D-fructose 1,6-bisphosphate	mutant R164A, Hill coefficient 2.1, pH 8.0, 28°C	Synechocystis sp.
2.6	-	D-fructose 1,6-bisphosphate	mutant R314A, Hill coefficient 1.8, pH 8.0, 28°C	Synechocystis sp.
5.2	-	D-fructose 1,6-bisphosphate	mutant F309A, Hill coefficient 1.8, pH 8.0, 28°C	Synechocystis sp.
5.2	-	D-fructose 1,6-bisphosphate	mutant N213A, Hill coefficient 1.8, pH 8.0, 28°C	Synechocystis sp.
5.2	-	D-fructose 1,6-bisphosphate	mutant R307A, Hill coefficient 1.6, pH 8.0, 28°C	Synechocystis sp.
7.1	-	D-fructose 1,6-bisphosphate	mutant R178A, Hill coefficient 1.1, pH 8.0, 28°C	Synechocystis sp.
10.5	-	D-fructose 1,6-bisphosphate	wild-type, Hill coefficient 1.9, pH 8.0, 28°C	Synechocystis sp.
21.3	-	D-fructose 1,6-bisphosphate	mutant K29A, Hill coefficient 2.0, pH 8.0, 28°C	Synechocystis sp.

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.015	-	mutant Y131A, pH 8.0, 28°C	Synechocystis sp.	AMP
0.0214	-	mutant R176A, pH 8.0, 28°C	Synechocystis sp.	AMP
0.027	-	mutant R178A, pH 8.0, 28°C	Synechocystis sp.	AMP
0.028	-	mutant K29A, pH 8.0, 28°C	Synechocystis sp.	AMP
0.034	-	wild-type, pH 8.0, 28°C	Synechocystis sp.	AMP
0.21	-	mutant N213A, pH 8.0, 28°C	Synechocystis sp.	AMP
0.225	-	mutant D200A, pH 8.0, 28°C	Synechocystis sp.	AMP
1.23	-	mutant D198H, pH 8.0, 28°C	Synechocystis sp.	AMP
1.241	-	mutant T102A, pH 8.0, 28°C	Synechocystis sp.	AMP
2.099	-	mutant K134A, pH 8.0, 28°C	Synechocystis sp.	AMP

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.21	-	D-fructose 1,6-bisphosphate	mutant R176A, pH 8.0, 28°C	Synechocystis sp.
0.48	-	D-fructose 1,6-bisphosphate	mutant D200A, pH 8.0, 28°C	Synechocystis sp.
3.6	-	D-fructose 1,6-bisphosphate	mutant K134A, pH 8.0, 28°C	Synechocystis sp.
4.7	-	D-fructose 1,6-bisphosphate	mutant Y131A, pH 8.0, 28°C	Synechocystis sp.
4.8	-	D-fructose 1,6-bisphosphate	mutant D198H, pH 8.0, 28°C	Synechocystis sp.
6.8	-	D-fructose 1,6-bisphosphate	mutant T102A, pH 8.0, 28°C	Synechocystis sp.
12	-	D-fructose 1,6-bisphosphate	mutant H215A, pH 8.0, 28°C	Synechocystis sp.
12	-	D-fructose 1,6-bisphosphate	mutant R178A, pH 8.0, 28°C	Synechocystis sp.
22	-	D-fructose 1,6-bisphosphate	mutant R164A, pH 8.0, 28°C	Synechocystis sp.
22	-	D-fructose 1,6-bisphosphate	mutant R307A, pH 8.0, 28°C	Synechocystis sp.
25	-	D-fructose 1,6-bisphosphate	mutant F309A, pH 8.0, 28°C	Synechocystis sp.
32	-	D-fructose 1,6-bisphosphate	mutant R314A, pH 8.0, 28°C	Synechocystis sp.
61	-	D-fructose 1,6-bisphosphate	mutant N213A, pH 8.0, 28°C	Synechocystis sp.
130	-	D-fructose 1,6-bisphosphate	wild-type, pH 8.0, 28°C	Synechocystis sp.
140	-	D-fructose 1,6-bisphosphate	mutant K29A, pH 8.0, 28°C	Synechocystis sp.