Cloned (Comment) | Organism |
---|---|
expression of SecS as C-terminally His6-tagged enzyme in Escherichia coli strain Rosetta2(DE3) | Mus musculus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant C-terminally His6-tagged SecS, apoenzyme or in complex with phosphate or iodide, sitting-drop vapor diffusion at 20°C, 14 mg/ml protein in 10 mM HEPES-NaOH, pH 7.5, 500 mM NaCl, 2 mM DTT, is mixed with an equal volume of reservoir solution containing 11% v/v ethylene glycol without other buffer components, 2 days, cryoprotection by crystal soaking in 100 mM HEPES-NaOH, pH 7.5, 250 mM NaCl, 1 mM dithiothreitol, and 35% v/v ethylene glycol, X-ray diffraction structure determination and analysis at 1.65-2.38 A resolution | Mus musculus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
20000 | - |
about, gel filtration | Mus musculus |
49000 | - |
4 * 49000, SDS-PAGE | Mus musculus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-seryl-tRNASec + selenophosphate | Mus musculus | - |
L-selenocysteinyl-tRNASec + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His6-tagged SecS from Escherichia coli strain Rosetta2(DE3) by nickel affinity chromatography and gel filtration to over 95% purity | Mus musculus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
L-seryl-tRNASec + selenophosphate = L-selenocysteinyl-tRNASec + phosphate | substrate binding and reaction mechanism | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-seryl-tRNASec + selenophosphate | - |
Mus musculus | L-selenocysteinyl-tRNASec + phosphate | - |
? | |
L-seryl-tRNASec + selenophosphate | the phosphate loop accommodates the gamma-phosphate moiety of O-phospho-L-seryltRNA [Ser]Sec and, after phosphate elimination, binds selenophosphate to initiate attack on the proposed aminoacrylyltRNA [Ser]Sec intermediate. Binding of phosphate triggers disorder-order transition in an active site loop | Mus musculus | L-selenocysteinyl-tRNASec + phosphate | - |
? | |
additional information | SecS does not act on free O-phospho-L-serine | Mus musculus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | two SecS monomers interact intimately and together build up two identical active sites around PLP in a Schiff-base linkage with Lys284. Two SecS dimers further associate to form a homotetramer. The N-terminus, which mediates tetramer formation, and a large insertion that remodels the active site set SecS aside from other members of the family | Mus musculus |
tetramer | 4 * 49000, SDS-PAGE | Mus musculus |
Synonyms | Comment | Organism |
---|---|---|
More | SecS is a member of the fold type I family of PLP-dependent enzymes with distinct structural elements | Mus musculus |
SecS | - |
Mus musculus |
Selenocysteine synthase | - |
Mus musculus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | SecS is a a PLP-dependent transferase. The PLP cofactor iIs tightly anchored by non-canonical contacts to both protomers of a close dimer | Mus musculus |