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Literature summary for 5.5.1.9 extracted from
- Plant cyclopropylsterol-cycloisomerase: key amino acids affecting activity and substrate specificity (2014), Biochem. J., 459, 289-299.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed as a Flag-tagged fusion protein | Arabidopsis thaliana |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D260E | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 92%, wild-type: 100%. Relative activity: 135% (wild-type: 100%). Km (cycloeucalenol) increased compared to wild-type. Mutant shows 6% activity with 24-methylene-24-dihydro-cycloartenol (wild-type 0%) | Arabidopsis thaliana |
| D260H | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 61%, wild-type: 100%. Relative activity: 5.2% (wild-type: 100%). Km (cycloeucalenol) decreased compared to wild-type | Arabidopsis thaliana |
| D260V | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 0%, wild-type: 100%. Relative activity: below 1.5% (wild-type: 100%) | Arabidopsis thaliana |
| D260Y | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 22%, wild-type: 100%. Relative activity: 0.7% (wild-type: 100%) | Arabidopsis thaliana |
| E29D | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 65%, wild-type: 100%. Relative activity: 3.8% (wild-type: 100%). Km (cycloeucalenol) decreased compared to wild-type | Arabidopsis thaliana |
| E29H | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 33%, wild-type: 100%. Relative activity: 3.5% (wild-type: 100%). Km (cycloeucalenol) decreased compared to wild-type | Arabidopsis thaliana |
| E29K | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 1.7%, wild-type: 100%. Relative activity: below 0.5% (wild-type: 100%) | Arabidopsis thaliana |
| E29K | in vivo conversion of cycloeucalenol into ergosterol and in vitro CPI activity in Saccharomyces cerevisiae erg7 transformants carrying Arabidopsis thaliana mutant CPI: 1.7% relative ergosterol synthesis (wild-type: 100%), below 0.5% in vitro microsomal CPI activity (wild-type: 100%) | Arabidopsis thaliana |
| E29V | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 0%, wild-type: 100%. Relative activity: below 0.5% (wild-type: 100%) | Arabidopsis thaliana |
| E54V | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 94%, wild-type: 100% | Arabidopsis thaliana |
| E58V | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 91%, wild-type: 100% | Arabidopsis thaliana |
| E60V | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 83%, wild-type: 100% | Arabidopsis thaliana |
| F111L | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 88%, wild-type: 100%. Relative activity: 31% (wild-type: 100%). Km (cycloeucalenol) increased compared to wild-type | Arabidopsis thaliana |
| F116L | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 37%, wild-type: 100%. Relative activity: 4.7% (wild-type: 100%) | Arabidopsis thaliana |
| F117L | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 89%, wild-type: 100%. Relative activity: 28% (wild-type: 100%). Km (cycloeucalenol) increased compared to wild-type | Arabidopsis thaliana |
| G108A | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 92%, wild-type: 100%. Relative activity: 90% (wild-type: 100%) | Arabidopsis thaliana |
| G108E | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 0%, wild-type: 100%. Relative activity: below 0.5% (wild-type: 100%). Km (cycloeucalenol) increased compared to wild-type | Arabidopsis thaliana |
| G108E | in vivo conversion of cycloeucalenol into ergosterol and in vitro CPI activity in Saccharomyces cerevisiae erg7 transformants carrying Arabidopsis thaliana mutant CPI: 0% relative ergosterol synthesis (wild-type: 100%), below 0.5% in vitro microsomal CPI activity (wild-type: 100%) | Arabidopsis thaliana |
| G108V | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 6%, wild-type: 100%. Relative activity: 1.7% (wild-type: 100%) | Arabidopsis thaliana |
| G28E | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 15%, wild-type: 100%. Relative activity: 1% (wild-type: 100%) | Arabidopsis thaliana |
| G28E | in vivo conversion of cycloeucalenol into ergosterol and in vitro CPI activity in Saccharomyces cerevisiae erg7 transformants carrying Arabidopsis thaliana mutant CPI: 15% relative ergosterol synthesis (wild-type: 100%), 1% in vitro microsomal CPI activity (wild-type: 100%) | Arabidopsis thaliana |
| G28V | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 46%, wild-type: 100%. Relative activity: 1.6% (wild-type: 100%) | Arabidopsis thaliana |
| H114L | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 85%, wild-type: 100%. Relative activity: 5.2% (wild-type: 100%) | Arabidopsis thaliana |
| N109A | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 76%, wild-type: 100%. Relative activity: 4.6% (wild-type: 100%) | Arabidopsis thaliana |
| T113A | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 69%, wild-type: 100%. Relative activity: 7.2% (wild-type: 100%). Km (cycloeucalenol) increased compared to wild-type | Arabidopsis thaliana |
| T113I | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 66%, wild-type: 100%. Relative activity: 3.1% (wild-type: 100%) | Arabidopsis thaliana |
| T113I | in vivo conversion of cycloeucalenol into ergosterol and in vitro CPI activity in Saccharomyces cerevisiae erg7 transformants carrying Arabidopsis thaliana mutant CPI: 66% relative ergosterol synthesis (wild-type: 100%), below 2.4% in vitro microsomal CPI activity (wild-type: 100%) | Arabidopsis thaliana |
| T113S | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 98%, wild-type: 100%. Relative activity: 73% (wild-type: 100%). Km (cycloeucalenol) increased compared to wild-type | Arabidopsis thaliana |
| W112E | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 77%, wild-type: 100%. Relative activity: 5.8% (wild-type: 100%) | Arabidopsis thaliana |
| W112K | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 47%, wild-type: 100%. Relative activity: 1.8% (wild-type: 100%) | Arabidopsis thaliana |
| W112L | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 86%, wild-type: 100%. Relative activity: 49% (wild-type: 100%). Km (cycloeucalenol) decreased compared to wild-type. Mutant shows 69% activity with 24-methylene-24-dihydro-cycloartenol (wild-type 0%), 10% activity with cycloartenol (wild-type 0%). 32% activity with cycloartanol (wild-type 0%), 15% activity with 4beta-methyl-24-dihydro-cycloeucalenol (wild-type 0%) | Arabidopsis thaliana |
| Y110L | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 64%, wild-type: 100%. Relative activity: 2.8% (wild-type: 100%) | Arabidopsis thaliana |
| Y115L | cycloeucalenol conversion into ergosterol in Saccharomyces cerevisiae erg7 transformants carrying mutagenized AtCPI, relative ergosterol synthesis: 83%, wild-type: 100%. Relative activity: 4.3% (wild-type: 100%) | Arabidopsis thaliana |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.077 | - |
cycloeucalenol | mutant E29D, 30°C, pH not specified in the publication, Vmax: 0.24 nmol/h/mg | Arabidopsis thaliana |  |
| 0.078 | - |
cycloeucalenol | mutant D260H, 30°C, pH not specified in the publication, Vmax: 0.35 nmol/h/mg | Arabidopsis thaliana | |
| 0.093 | - |
cycloeucalenol | mutant E29H, 30°C, pH not specified in the publication, Vmax: 0.33 nmol/h/mg | Arabidopsis thaliana | |
| 0.135 | - |
cycloeucalenol | mutant W112L, 30°C, pH not specified in the publication, Vmax: 14.7 nmol/h/mg | Arabidopsis thaliana | |
| 0.167 | - |
cycloeucalenol | wild-type, 30°C, pH not specified in the publication, Vmax: 22 nmol/h/mg | Arabidopsis thaliana | |
| 0.377 | - |
cycloeucalenol | mutant F111L, 30°C, pH not specified in the publication, Vmax: 4.7 nmol/h/mg | Arabidopsis thaliana | |
| 0.409 | - |
cycloeucalenol | mutant D260E, 30°C, pH not specified in the publication, Vmax: 44 nmol/h/mg | Arabidopsis thaliana | |
| 0.496 | - |
cycloeucalenol | mutant T113S, 30°C, pH not specified in the publication, Vmax: 26 nmol/h/mg | Arabidopsis thaliana | |
| 0.52 | - |
cycloeucalenol | mutant F117L, 30°C, pH not specified in the publication, Vmax: 5.3 nmol/h/mg | Arabidopsis thaliana | |
| 0.622 | - |
cycloeucalenol | mutant G108A, 30°C, pH not specified in the publication, Vmax: 11.4 nmol/h/mg | Arabidopsis thaliana | |
| 1.53 | - |
cycloeucalenol | mutant T113A, 30°C, pH not specified in the publication, Vmax: 4.9 nmol/h/mg | Arabidopsis thaliana | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Arabidopsis thaliana | - |
- |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| cycloeucalenol | - |
Arabidopsis thaliana | obtusifoliol | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Atcpi | - |
Arabidopsis thaliana |
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Release 2023.1 (January 2023)
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