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Literature summary for 2.3.1.297 extracted from
- Substrate specificity, kinetic properties and inhibition by fumonisin B1 of ceramide synthase isoforms from Arabidopsis (2016), Biochem. J., 473, 593-603 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene LAG1, recombinant expression of codon-optimized LAG1 as N-terminally FLAG-tagged enzyme in Saccharomyces cerevisiae membranes | Arabidopsis thaliana |
| gene LAG3, recombinant expression of codon-optimized LAG3 as N-terminally FLAG-tagged enzyme in Saccharomyces cerevisiae membranes | Arabidopsis thaliana |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | complementation of the growth defect of DELTAlag1/DELTAlac1 yeast deletion mutant by recombinant GST-/FLAG-tagged LOH1. Upon isolation of a microsomal membrane fraction from yeast expressing the construct, LOH1 shows significant activity by an in vitro ceramide synthase assay | Arabidopsis thaliana |
| additional information | complementation of the growth defect of DELTAlag1/DELTAlac1 yeast deletion mutant by recombinant GST-/FLAG-tagged LOH3. Upon isolation of a microsomal membrane fraction from yeast expressing the construct, LOH3 shows significant activity by an in vitro ceramide synthase assay | Arabidopsis thaliana |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| fumonisin B1 | mixed mode of inhibition with respect to the laong-chain base; mixed mode of inhibition with respect to the long-chain base | Arabidopsis thaliana |  |
| additional information | LOH1 is inhibited by most or all divalent cations tested; LOH3 is inhibited by most or all divalent cations tested | Arabidopsis thaliana |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Arabidopsis thaliana | |
| 0.0069 | - |
phytosphingosine | pH 7.5, 30°C, LOH1 | Arabidopsis thaliana | |
| 0.023 | - |
phytosphingosine | pH 7.5, 30°C, LOH3 | Arabidopsis thaliana | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| microsome | - |
Arabidopsis thaliana | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| additional information | LOH1 is inhibited by most or all divalent cations tested | Arabidopsis thaliana | |
| additional information | LOH3 is inhibited by most or all divalent cations tested | Arabidopsis thaliana |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| hexacosanoyl-CoA + phytosphingosine | Arabidopsis thaliana | - |
CoA + N-hexacosanoylphytosphingosine | - |
? | |
| lignoceroyl-CoA + phytosphingosine | Arabidopsis thaliana | - |
N-lignoceroylphytosphinganine + CoA | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Arabidopsis thaliana | Q6NQI8 | - |
- |
| Arabidopsis thaliana | Q9LDF2 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| acyl-CoA + sphinganine | very low activity with sphinganine | Arabidopsis thaliana | N-acylsphinganine + CoA | - |
? | |
| hexacosanoyl-CoA + phytosphingosine | - |
Arabidopsis thaliana | CoA + N-hexacosanoylphytosphingosine | - |
? | |
| lignoceroyl-CoA + phytosphingosine | - |
Arabidopsis thaliana | N-lignoceroylphytosphinganine + CoA | - |
? | |
| lignoceroyl-CoA + phytosphingosine | high activity | Arabidopsis thaliana | N-lignoceroylphytosphinganine + CoA | - |
? | |
| additional information | specificity of three different isoforms of ceramide synthase, denoted LOH1, 2 and 3, for a range of long-chain base (LCB) and acyl-CoA substrates. The contribution of each of these isoforms to the synthesis of ceramide is investigated by in vitro ceramide synthase assays, overview. The plant LCB phytosphingosine is efficiently used by the LOH1 and 3 isoforms, with LOH1 having the lowest Km for the LCB substrate of the three isoforms. In contrast, sphinganine is used efficiently only by the LOH2 isoform. Acyl-CoA specificity is also distinguished between the three isoforms with LOH2 being almost completely specific for palmitoyl-CoA, while the LOH1 isoform shows greatest activity with lignoceroyl- and hexacosanoyl-CoA. Unsaturated acyl-CoAs are not used efficiently by any isoform while unsaturated LCB substrates are preferred by LOH2 and 3. LOH3 has at least twice as much activity with t18:1 substrates as LOH1. Both LOH1 and LOH3 demonstrate a strong preference for very long chain acyl-CoAs (>C18) although LOH1 has the greatest activity toward 24 and 26 carbon acyl-CoAs, while LOH3 shows little preference for acyl-CoAs between 20 and 26 carbons in length | Arabidopsis thaliana | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ceramide synthase | - |
Arabidopsis thaliana |
| LAG1 longevity assurance homolog 1 | - |
Arabidopsis thaliana |
| LAG1 longevity assurance homolog 3 | - |
Arabidopsis thaliana |
| LAG11 | - |
Arabidopsis thaliana |
| LAG13 | - |
Arabidopsis thaliana |
| LOH1 | - |
Arabidopsis thaliana |
| LOH3 | - |
Arabidopsis thaliana |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Arabidopsis thaliana |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Arabidopsis thaliana |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.000003 | - |
fumonisin B1 | pH 7.5, 30°C, LOH1 | Arabidopsis thaliana | |
| 0.000784 | - |
fumonisin B1 | pH 7.5, 30°C, LOH3 | Arabidopsis thaliana | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| physiological function | distinct roles and modes of regulation for each of the ceramide synthases in Arabidopsis thaliana sphingolipid metabolism. Ceramides are organizing components of sphingolipids in the eukaryotic cell. Three ceramide synthase isoforms are found in Arabidopsis thaliana each with specific substrate preferences and sensitivity to inhibitors and activators, overview. Isozymes LOH1 and LOH3 are responsible for the synthesis of ceramides with trihydroxy long-chain bases and very long chain fatty acids (VLCFA) | Arabidopsis thaliana |
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