Literature summary extracted from
Hu, X.; Murata, L.B.; Weichsel, A.; Brailey, J.L.; Roberts, S.A.; Nighorn, A.; Montfort, W.R.
Allostery in recombinant soluble guanylyl cyclase from Manduca sexta (2008), J. Biol. Chem., 283, 20968-20977.
Activating Compound
EC Number |
Activating Compound |
Comment |
Organism |
Structure |
---|
4.6.1.2 |
additional information |
for maximal activity, msGC required both NO and YC-1 |
Manduca sexta |
|
4.6.1.2 |
NO |
170fold activation, binding of NO leads to a transient six-coordinate intermediate, followed by release of the proximal histidine to yield a five-coordinate nitrosyl complex. Hallmark of sGC activation by NO is the release of beta1 His105 from the heme, leading to allosteric stimulation of cyclase activity |
Manduca sexta |
|
4.6.1.2 |
YC-1 |
affects binding of NO and CO to the enzyme, it reduces the NO and CO off-rates for the 100 kDa N-terminal heterodimeric fragment and increases the CO affinity by 50fold, overview |
Manduca sexta |
|
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
4.6.1.2 |
expression of His-tagged heterodimeric full-length and N-terminal fragments of Manduca sexta sGC in Escherichia coli strains BL21(DE3) |
Manduca sexta |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
4.6.1.2 |
alpha1L211A |
site-directed mutagenesis of the alpha1 subunit |
Manduca sexta |
4.6.1.2 |
alpha1Y223A |
site-directed mutagenesis |
Manduca sexta |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
4.6.1.2 |
ATP |
inhibits the enzyme by 70% at 1 mM in presence of stoichiometric concentrations of NO |
Manduca sexta |
|
KM Value [mM]
EC Number |
KM Value [mM] |
KM Value Maximum [mM] |
Substrate |
Comment |
Organism |
Structure |
---|
4.6.1.2 |
additional information |
- |
additional information |
kinetics of ligand binding |
Manduca sexta |
|
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
4.6.1.2 |
CO |
competitive binding, off rates are reduced by YC-1, but YC-1 stimulation of CO binding is unaltered for the heme-intact portion of both mutated proteins, CO release from msGC-NT1-CO is measured in a stopped-flow device by replacement with NO, which binds more quickly and tightly to the enzyme, overview |
Manduca sexta |
|
4.6.1.2 |
Mg2+ |
- |
Manduca sexta |
|
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
4.6.1.2 |
GTP |
Manduca sexta |
- |
3',5'-cyclic GMP + diphosphate |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
4.6.1.2 |
Manduca sexta |
Q8IT60 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
4.6.1.2 |
recombinant His-tagged heterodimeric full-length and N-terminal fragments of sGC from Escherichia coli by nickel affinity chromatography and gel filtration |
Manduca sexta |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
4.6.1.2 |
GTP |
- |
Manduca sexta |
3',5'-cyclic GMP + diphosphate |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
4.6.1.2 |
More |
structure molecular modeling of N-terminal domains of GC alpha1 and beta1 subunits |
Manduca sexta |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
4.6.1.2 |
sGC |
- |
Manduca sexta |
4.6.1.2 |
soluble guanylyl cyclase |
- |
Manduca sexta |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
4.6.1.2 |
22 |
- |
assay at room temperature |
Manduca sexta |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
4.6.1.2 |
7.4 |
7.5 |
assay at |
Manduca sexta |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
4.6.1.2 |
heme |
hallmark of sGC activation by NO is the release of beta1 His105 from the heme, leading to allosteric stimulation of cyclase activity |
Manduca sexta |
|