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Literature summary for 2.4.3.1 extracted from
- The sialyltransferase 'sialylmotif' participates in binding the donor substrate CMP-NeuAc (1995), J. Biol. Chem., 270, 1497-1500.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of wild-type and mutants in COS-1 cells | Rattus norvegicus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D219A | site-directed mutagenesis, mutant of the conserved sialyl-motif contained by all members of the sialyltransferase family, 10.8fold increased Km for CMP-NeuAc, only slightly altered Km for the acceptor substrate | Rattus norvegicus |
| G221A | site-directed mutagenesis, mutant of the conserved sialyl-motif contained by all members of the sialyltransferase family, 1.6fold increased Km for CMP-NeuAc, only slightly altered Km for the acceptor substrate | Rattus norvegicus |
| K223A | site-directed mutagenesis, mutant of the conserved sialyl-motif contained by all members of the sialyltransferase family, 6.6fold increased Km for CMP-NeuAc, only slightly altered Km for the acceptor substrate | Rattus norvegicus |
| L190A | site-directed mutagenesis, mutant of the conserved sialyl-motif contained by all members of the sialyltransferase family, 11.9fold increased Km for CMP-NeuAc, only slightly altered Km for the acceptor substrate | Rattus norvegicus |
| T224A | site-directed mutagenesis, mutant of the conserved sialyl-motif contained by all members of the sialyltransferase family, 1.3fold increased Km for CMP-NeuAc, only slightly altered Km for the acceptor substrate | Rattus norvegicus |
| T225A | site-directed mutagenesis, mutant of the conserved sialyl-motif contained by all members of the sialyltransferase family, 3.2fold increased Km for CMP-NeuAc, only slightly altered Km for the acceptor substrate | Rattus norvegicus |
| V184A | site-directed mutagenesis, mutant of the conserved sialyl-motif contained by all members of the sialyltransferase family, 6.8fold increased Km for CMP-NeuAc, only slightly altered Km for the acceptor substrate | Rattus norvegicus |
| V220A | site-directed mutagenesis, mutant of the conserved sialyl-motif contained by all members of the sialyltransferase family, 6.0fold increased Km for CMP-NeuAc, only slightly altered Km for the acceptor substrate | Rattus norvegicus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| CDP | competitive; wild-type enzyme and mutant V220A | Rattus norvegicus |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.05 | - |
CMP-N-acetylneuraminate | wild-type enzyme | Rattus norvegicus | |
| 0.05 | - |
CMP-N-acetylneuraminate | with asialo-alpha1-acid glycoprotein | Rattus norvegicus | |
| 0.33 | - |
asialo-alpha1-acid glycoprotein | wild-type enzyme | Rattus norvegicus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Rattus norvegicus | P13721 | - |
- |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| CMP-N-acetyl-beta-neuraminate + beta-D-galactosyl-R = CMP + N-acetyl-alpha-neuraminyl-(2->6)-beta-D-galactosyl-R | Val220 participates in binding the common cytidine moiety in substrate CMP-NeuAc and inhibitor CDP | Rattus norvegicus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| CMP-N-acetylneuraminate + asialo-alpha1-acid glycoprotein | - |
Rattus norvegicus | CMP + alpha1-acid glycoprotein | - |
? | |
| CMP-N-acetylneuraminate + asialo-orosomucoid | i.e. asialo-alpha1 acid glycoprotein | Rattus norvegicus | CMP + orosomucoid | - |
? | |
| CMP-N-acetylneuraminate + beta-D-galactosyl-1,4-N-acetyl-beta-D-glucosaminyl-R | sialic acid is linked to C-6 of the galactose residue | Rattus norvegicus | CMP + alpha-N-acetylneuraminyl-2,6-beta-D-galactosyl-1,4-N-acetyl-beta-D-glucosaminyl-R | - |
? | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Rattus norvegicus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | - |
assay at | Rattus norvegicus |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0047 | - |
CDP | wild-type enzyme | Rattus norvegicus | |
| 0.016 | - |
CDP | mutant V220A | Rattus norvegicus | |
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