Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Sphingomonas paucimobilis |
Crystallization (Comment) | Organism |
---|---|
purified His-tagged recombinant wild-type and mutant enzymes in complex with cofactor and substrates, wild-type enzyme from 10 mM Tris, pH 7.5, 150 mM NaCl, and 0.025 mM or 0.250 mM pyridoxal 5'-phosphate, different conditions for the mutants, overview. Mass spectroscopic structure analysis, overview | Sphingomonas paucimobilis |
Protein Variants | Comment | Organism |
---|---|---|
N100C | site-directed mutagenesis, the mutation mimics the wild-type human enzyme and is fully active, crystal structure analysis | Sphingomonas paucimobilis |
N100W | site-directed mutagenesis, the mutation mimics the mutation in the human enzyme causing hereditary sensory autonomic neuropathy type 1, the mutant shows reduced activity compared to the wild-type enzyme. The mutation affects the chemistry of the pyridoxal 5'-phosphate, crystal structure analysis | Sphingomonas paucimobilis |
N100Y | site-directed mutagenesis, N100Y is less able to stabilize a quinonoid intermediate, the mutation mimics the mutation in the human enzyme causing hereditary sensory autonomic neuropathy type 1, the mutant shows reduced activity compared to the wild-type enzyme. The mutation affects the chemistry of the pyridoxal 5'-phosphate. The L-Ser external aldimine structure N100Y reveals significant differences that hinder the movement of a catalytically important Arg378 residue into the active site, crystal structure analysis | Sphingomonas paucimobilis |
R378A | site-directed mutagenesis, crystal structure analysis, the mutant is less able to stabilize a quinonoid intermediate | Sphingomonas paucimobilis |
R378N | site-directed mutagenesis, crystal structure analysis | Sphingomonas paucimobilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.019 | - |
palmitoyl-CoA | pH 7.5, 37°C, recombinant mutant N100W | Sphingomonas paucimobilis | |
0.031 | - |
palmitoyl-CoA | pH 7.5, 37°C, recombinant mutant N100Y | Sphingomonas paucimobilis | |
0.031 | - |
palmitoyl-CoA | pH 7.5, 37°C, recombinant mutant R378N | Sphingomonas paucimobilis | |
0.035 | - |
palmitoyl-CoA | pH 7.5, 37°C, recombinant wild-type enzyme | Sphingomonas paucimobilis | |
0.039 | - |
palmitoyl-CoA | pH 7.5, 37°C, recombinant mutant R378A | Sphingomonas paucimobilis | |
0.06 | - |
palmitoyl-CoA | pH 7.5, 37°C, recombinant mutant N100C | Sphingomonas paucimobilis | |
1.4 | - |
L-serine | pH 7.5, 37°C, recombinant wild-type enzyme | Sphingomonas paucimobilis | |
1.6 | - |
L-serine | pH 7.5, 37°C, recombinant mutant N100W | Sphingomonas paucimobilis | |
2.4 | - |
L-serine | pH 7.5, 37°C, recombinant mutant R378N | Sphingomonas paucimobilis | |
2.5 | - |
L-serine | pH 7.5, 37°C, recombinant mutant N100Y | Sphingomonas paucimobilis | |
3.8 | - |
L-serine | pH 7.5, 37°C, recombinant mutant R378A | Sphingomonas paucimobilis | |
7 | - |
L-serine | pH 7.5, 37°C, recombinant mutant N100C | Sphingomonas paucimobilis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | bound | Homo sapiens | 16020 | - |
membrane | bound | Sphingomonas paucimobilis | 16020 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
palmitoyl-CoA + L-serine | Homo sapiens | - |
CoA + 3-dehydro-D-sphinganine + CO2 | - |
? | |
palmitoyl-CoA + L-serine | Sphingomonas paucimobilis | - |
CoA + 3-dehydro-D-sphinganine + CO2 | - |
? | |
palmitoyl-CoA + L-serine | Sphingomonas paucimobilis EY2395 | - |
CoA + 3-dehydro-D-sphinganine + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Sphingomonas paucimobilis | Q93UV0 | - |
- |
Sphingomonas paucimobilis EY2395 | Q93UV0 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Sphingomonas paucimobilis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
palmitoyl-CoA + L-serine = CoA + 3-dehydro-D-sphinganine + CO2 | the enzyme forms an external aldimine intermediate, as well as dunathan and quinoid intermediates, and followed by external beta-keto acid intermediate, and finally product quinoid and product external aldimine intermediates, reaction cycle, overview | Sphingomonas paucimobilis | |
palmitoyl-CoA + L-serine = CoA + 3-dehydro-D-sphinganine + CO2 | the enzyme forms an external aldimine intermediate, as well as dunathan and quinoid intermediates, followed by external beta-keto acid intermediate, and finally product quinoid and product external aldimine intermediates, reaction cycle, overview | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
palmitoyl-CoA + L-serine | - |
Homo sapiens | CoA + 3-dehydro-D-sphinganine + CO2 | - |
? | |
palmitoyl-CoA + L-serine | - |
Sphingomonas paucimobilis | CoA + 3-dehydro-D-sphinganine + CO2 | - |
? | |
palmitoyl-CoA + L-serine | - |
Sphingomonas paucimobilis EY2395 | CoA + 3-dehydro-D-sphinganine + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | structure comparison with the SPT from Sphingomonas paucimobilis, overview | Homo sapiens |
homodimer | structure comparison with the human SPT, overview | Sphingomonas paucimobilis |
Synonyms | Comment | Organism |
---|---|---|
serine palmitoyltransferase | - |
Homo sapiens |
serine palmitoyltransferase | - |
Sphingomonas paucimobilis |
SPT | - |
Homo sapiens |
SPT | - |
Sphingomonas paucimobilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Sphingomonas paucimobilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.005 | - |
L-serine | recombinant mutant N100Y | Sphingomonas paucimobilis | |
0.005 | - |
palmitoyl-CoA | recombinant mutant N100Y | Sphingomonas paucimobilis | |
0.009 | - |
L-serine | recombinant mutant N100W | Sphingomonas paucimobilis | |
0.009 | - |
palmitoyl-CoA | recombinant mutant N100W | Sphingomonas paucimobilis | |
0.033 | - |
L-serine | recombinant mutant R378N | Sphingomonas paucimobilis | |
0.033 | - |
palmitoyl-CoA | recombinant mutant R378N | Sphingomonas paucimobilis | |
0.078 | - |
L-serine | recombinant mutant R378A | Sphingomonas paucimobilis | |
0.078 | - |
palmitoyl-CoA | recombinant mutant R378A | Sphingomonas paucimobilis | |
0.25 | - |
L-serine | recombinant mutant N100C | Sphingomonas paucimobilis | |
0.25 | - |
palmitoyl-CoA | recombinant mutant N100C | Sphingomonas paucimobilis | |
1.15 | - |
L-serine | recombinant wild-type enzyme | Sphingomonas paucimobilis | |
1.15 | - |
palmitoyl-CoA | recombinant wild-type enzyme | Sphingomonas paucimobilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Sphingomonas paucimobilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | dependent on | Homo sapiens | |
pyridoxal 5'-phosphate | dependent on, bound at the active site at the dimer interface | Sphingomonas paucimobilis |
General Information | Comment | Organism |
---|---|---|
malfunction | mutations in human SPT cause hereditary sensory autonomic neuropathy type 1, HSAN1, a disease characterized by loss of feeling in extremities and severe pain | Homo sapiens |
metabolism | SPT catalyzes the first and rate-limiting step of the sphingolipid biosynthetic pathway | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.002 | - |
L-serine | pH 7.5, 37°C, recombinant mutant N100Y | Sphingomonas paucimobilis | |
0.006 | - |
L-serine | pH 7.5, 37°C, recombinant mutant N100W | Sphingomonas paucimobilis | |
0.014 | - |
L-serine | pH 7.5, 37°C, recombinant mutant R378N | Sphingomonas paucimobilis | |
0.021 | - |
L-serine | pH 7.5, 37°C, recombinant mutant R378A | Sphingomonas paucimobilis | |
0.036 | - |
L-serine | pH 7.5, 37°C, recombinant mutant N100C | Sphingomonas paucimobilis | |
0.821 | - |
L-serine | pH 7.5, 37°C, recombinant wild-type enzyme | Sphingomonas paucimobilis |