Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | for maximal activity, msGC required both NO and YC-1 | Manduca sexta | |
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 | |
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 (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
alpha1L211A | site-directed mutagenesis of the alpha1 subunit | Manduca sexta |
alpha1Y223A | site-directed mutagenesis | Manduca sexta |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ATP | inhibits the enzyme by 70% at 1 mM in presence of stoichiometric concentrations of NO | Manduca sexta |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of ligand binding | Manduca sexta |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
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 | |
Mg2+ | - |
Manduca sexta |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP | Manduca sexta | - |
3',5'-cyclic GMP + diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Manduca sexta | Q8IT60 | - |
- |
Purification (Comment) | Organism |
---|---|
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP | - |
Manduca sexta | 3',5'-cyclic GMP + diphosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | structure molecular modeling of N-terminal domains of GC alpha1 and beta1 subunits | Manduca sexta |
Synonyms | Comment | Organism |
---|---|---|
sGC | - |
Manduca sexta |
soluble guanylyl cyclase | - |
Manduca sexta |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Manduca sexta |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | 7.5 | assay at | Manduca sexta |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
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 |