Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain DH5alpha | Bradyrhizobium japonicum |
DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain DH5alpha | Rhodopseudomonas palustris |
DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain DH5alpha | Streptomyces peucetius |
DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain DH5alpha | Zymomonas mobilis |
DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain DH5alpha | Methylococcus capsulatus |
DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain DH5alpha | Tetrahymena thermophila |
DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain DH5alpha | Alicyclobacillus acidocaldarius |
gene shc, DNA and amino acid sequence determination, expression in Escherichia coli | Alicyclobacillus acidocaldarius |
Protein Variants | Comment | Organism |
---|---|---|
C435S/D374I/D374V/H451F | inactive mutant | Alicyclobacillus acidocaldarius |
C435S/D374I/D374V/H451F | site-directed mutagenesis, inactive mutant | Alicyclobacillus acidocaldarius |
D376E | inactive mutant | Alicyclobacillus acidocaldarius |
D376E | site-directed mutagenesis, inactive mutant | Alicyclobacillus acidocaldarius |
D377C/D377N/Y612A | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
D377C/D377N/Y612A | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
D377E/D376Q/D376R/D377R/E45K/W406V/W417A/D377C | inactive mutant | Alicyclobacillus acidocaldarius |
D377E/D376Q/D376R/D377R/E45K/W406V/W417A/D377C | site-directed mutagenesis, inactive mutant | Alicyclobacillus acidocaldarius |
F365A | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
F365A | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
F601A | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
F601A | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
F605A | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
F605A | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
I261A | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
I261A | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
additional information | product patterns of mutant enzymes, detailed overview | Alicyclobacillus acidocaldarius |
Q262G/Q262A/P263G/P263A | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
Q262G/Q262A/P263G/P263A | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
V380E | inactive mutant | Alicyclobacillus acidocaldarius |
V380E | site-directed mutagenesis, inactive mutant | Alicyclobacillus acidocaldarius |
V381A/D376C | inactive mutant | Alicyclobacillus acidocaldarius |
V381A/D376C | site-directed mutagenesis, inactive mutant | Alicyclobacillus acidocaldarius |
W169F/W169H/W489A/F605K | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
W169F/W169H/W489A/F605K | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
Y420A | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
Y420A | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
Y606A/W23V/W495V/W522V/W533A/W591L/W78S/E35Q/E197Q/D530N/T378A | the mutant shows the same product pattern and activity as the wild-type | Alicyclobacillus acidocaldarius |
Y606A/W23V/W495V/W522V/W533A/W591L/W78S/E35Q/E197Q/D530N/T378A | site-directed mutagenesis, the mutant shows the same product pattern and activity as the wild-type | Alicyclobacillus acidocaldarius |
Y609A/Y612A/L607K | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
Y609A/Y612A/L607K | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
Y609F | the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
Y609F | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview | Alicyclobacillus acidocaldarius |
Y609F | site-directed mutagenesis, the mutant shows an altered product pattern compared to the wild-type enzyme, overview. The phenotype of Y609F mutein is contrarily described in two publications | Alicyclobacillus acidocaldarius |
Y612F/D376E/D376G/D377E/D377G/D377Q/E45A/E45D/F365W/T41A/E93A/R127Q/W133A/Y267A/F434A/F437A/W258L/D350N/D421N/D442N/H451R/D447N/D377N/D313N/E535Q/D374E | the mutant shows the same product pattern as the wild-type with less enzyme activity | Alicyclobacillus acidocaldarius |
Y612F/D376E/D376G/D377E/D377G/D377Q/E45A/E45D/F365W/T41A/E93A/R127Q/W133A/Y267A/F434A/F437A/W258L/D350N/D421N/D442N/H451R/D447N/D377N/D313N/E535Q/D374E | site-directed mutagenesis, the mutant shows the same product pattern as the wild-type with less enzyme activity | Alicyclobacillus acidocaldarius |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | 0.016 | squalene | pH 6.0, 60°C | Alicyclobacillus acidocaldarius |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
plasma membrane | enzyme SHC in vivo is a membrane-associated protein and can be solubilized from cell extracts by nonionic detergents, such as Triton X-100 or octylthioglucopyranoside. The enzyme is attached to the inner side of the cytoplasmic membrane by interactions of hydrophobic residues with the phospholipids. The membrane-binding part of the enzyme is a nonpolar region that is encircled by positive-charged amino acids enforcing the anchoring of the enzyme to the negatively charged surface of the phospholipid membrane | Alicyclobacillus acidocaldarius | 5886 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
71600 | - |
x * 71600, SDS-PAGE | Alicyclobacillus acidocaldarius |
71600 | - |
2 * 71600, about, sequence calculation | Alicyclobacillus acidocaldarius |
72000 | - |
x * 72000, SDS-PAGE | Tetrahymena thermophila |
72300 | - |
x * 72300, SDS-PAGE | Rhodopseudomonas palustris |
74100 | - |
x * 74100, SDS-PAGE | Streptomyces peucetius |
74100 | - |
x * 74100, SDS-PAGE | Zymomonas mobilis |
74100 | - |
x * 74100, SDS-PAGE | Methylococcus capsulatus |
76300 | - |
x * 76300, SDS-PAGE | Bradyrhizobium japonicum |
76300 | - |
x * 76300, about, sequence calculation | Bradyrhizobium japonicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Alicyclobacillus acidocaldarius | product pattern of alternative substrates, overview | ? | - |
? | |
squalene | Bradyrhizobium japonicum | - |
hop-22(29)-ene | - |
? | |
squalene | Rhodopseudomonas palustris | - |
hop-22(29)-ene | - |
? | |
squalene | Streptomyces peucetius | - |
hop-22(29)-ene | - |
? | |
squalene | Zymomonas mobilis | - |
hop-22(29)-ene | - |
? | |
squalene | Methylococcus capsulatus | - |
hop-22(29)-ene | - |
? | |
squalene | Tetrahymena thermophila | - |
hop-22(29)-ene | - |
? | |
squalene | Alicyclobacillus acidocaldarius | - |
hop-22(29)-ene | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Alicyclobacillus acidocaldarius | - |
formerly Bacillus acidocaldarius | - |
Alicyclobacillus acidocaldarius | P33247 | formerly Bacillus acidocaldarius, gene shc | - |
Bradyrhizobium japonicum | - |
- |
- |
Methylococcus capsulatus | - |
- |
- |
no activity in Methylococcus capsulatus | - |
- |
- |
Rhodopseudomonas palustris | - |
- |
- |
Streptomyces peucetius | - |
- |
- |
Tetrahymena thermophila | - |
- |
- |
Zymomonas mobilis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
native and/or recombinant enzyme, the enzyme in vivo is a membrane-associated protein and can be solubilized from cell extracts by nonionic detergents, such as Triton X-100 or octylthioglucopyranoside | Alicyclobacillus acidocaldarius |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
squalene = hop-22(29)-ene | catalytic mechanism, the initial reaction catalyzed is the protonation of the terminal double bond of squalene. The conserved DXDD motif of SHCs is essential for this protonation reaction, overview | Bradyrhizobium japonicum | |
squalene = hop-22(29)-ene | catalytic mechanism, the initial reaction catalyzed is the protonation of the terminal double bond of squalene. The conserved DXDD motif of SHCs is essential for this protonation reaction, overview | Rhodopseudomonas palustris | |
squalene = hop-22(29)-ene | catalytic mechanism, the initial reaction catalyzed is the protonation of the terminal double bond of squalene. The conserved DXDD motif of SHCs is essential for this protonation reaction, overview | Streptomyces peucetius | |
squalene = hop-22(29)-ene | catalytic mechanism, the initial reaction catalyzed is the protonation of the terminal double bond of squalene. The conserved DXDD motif of SHCs is essential for this protonation reaction, overview | Zymomonas mobilis | |
squalene = hop-22(29)-ene | catalytic mechanism, the initial reaction catalyzed is the protonation of the terminal double bond of squalene. The conserved DXDD motif of SHCs is essential for this protonation reaction, overview | Methylococcus capsulatus | |
squalene = hop-22(29)-ene | catalytic mechanism, the initial reaction catalyzed is the protonation of the terminal double bond of squalene. The conserved DXDD motif of SHCs is essential for this protonation reaction, overview | Tetrahymena thermophila | |
squalene = hop-22(29)-ene | catalytic mechanism, the initial reaction catalyzed is the protonation of the terminal double bond of squalene. The conserved DXDD motif of SHCs is essential for this protonation reaction. In Alicyclobacillus acidocaldarius SHC, Asp376 of this motif is hydrogen bonded to His451, and an additional hydrogen bond exists to an ordered water molecule, which connects D376 to the hydroxyl group of the Y495 side chain and thus further enhances its acidity. The carboxyl groups of Asp374 and Asp377 accommodate the positive charge of the D376-H451 pair prior to proton transfer. After proton transfer to the 2,3-double bond of squalene, the D376-H451 pair loses its charge, leaving the remaining negative charge on the D374-D377 pair for stabilization of the initial cationic intermediates (24, 101). Reprotonation of D376 occurs through a water molecule bound to Y495-OH, which can transfer protons from disordered water in the solvent-accessible upper cavity of SHC | Alicyclobacillus acidocaldarius | |
squalene = hop-22(29)-ene | overall mechanism of the polycyclization reaction of SHCs and structures of squalene cyclization products, overview | Bradyrhizobium japonicum | |
squalene = hop-22(29)-ene | overall mechanism of the polycyclization reaction of SHCs and structures of squalene cyclization products, overview | Alicyclobacillus acidocaldarius |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | substrate specificity, overview | Alicyclobacillus acidocaldarius | ? | - |
? | |
additional information | substrate specificity, detailed overview | Streptomyces peucetius | ? | - |
? | |
additional information | substrate specificity, detailed overview | Zymomonas mobilis | ? | - |
? | |
additional information | the enzyme also catalyzes 2,3-oxidosqualene cyclization, but no tetrahymanol formation. Substrate specificity, detailed overview | Bradyrhizobium japonicum | ? | - |
? | |
additional information | the enzyme also catalyzes 2,3-oxidosqualene cyclization, but no tetrahymanol formation. Substrate specificity, detailed overview | Alicyclobacillus acidocaldarius | ? | - |
? | |
additional information | the enzyme also catalyzes 2,3-oxidosqualene cyclization, substrate specificity, detailed overview | Tetrahymena thermophila | ? | - |
? | |
additional information | the enzyme does not catalyze 2,3-oxidosqualene cyclization nor tetrahymanol formation. Substrate specificity, detailed overview | Methylococcus capsulatus | ? | - |
? | |
additional information | the enzyme does not catalyze tetrahymanol formation. Substrate specificity, detailed overview | Rhodopseudomonas palustris | ? | - |
? | |
additional information | product pattern of alternative substrates, overview | Alicyclobacillus acidocaldarius | ? | - |
? | |
squalene | - |
Bradyrhizobium japonicum | hop-22(29)-ene | - |
? | |
squalene | - |
Rhodopseudomonas palustris | hop-22(29)-ene | - |
? | |
squalene | - |
Streptomyces peucetius | hop-22(29)-ene | - |
? | |
squalene | - |
Zymomonas mobilis | hop-22(29)-ene | - |
? | |
squalene | - |
Methylococcus capsulatus | hop-22(29)-ene | - |
? | |
squalene | - |
Tetrahymena thermophila | hop-22(29)-ene | - |
? | |
squalene | - |
Alicyclobacillus acidocaldarius | hop-22(29)-ene | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 72000, SDS-PAGE | Tetrahymena thermophila |
? | x * 71600, SDS-PAGE | Alicyclobacillus acidocaldarius |
? | x * 72300, SDS-PAGE | Rhodopseudomonas palustris |
? | x * 74100, SDS-PAGE | Streptomyces peucetius |
? | x * 74100, SDS-PAGE | Zymomonas mobilis |
? | x * 74100, SDS-PAGE | Methylococcus capsulatus |
? | x * 76300, SDS-PAGE | Bradyrhizobium japonicum |
? | x * 76300, about, sequence calculation | Bradyrhizobium japonicum |
homodimer | 2 * 71600, about, sequence calculation | Alicyclobacillus acidocaldarius |
More | dumbbell-shaped structure of chain A with a more structured beta-barrel structure in domain 1, active site structure, structure analysis, overview | Alicyclobacillus acidocaldarius |
More | each subunit consists of alpha-helical domains that build up a dumbbell-shaped structure. The first domain consists of a regular (alpha/alpha)6 barrel structure, whereas the second domain shows an alpha-barrel structure in a less periodic manner | Alicyclobacillus acidocaldarius |
Synonyms | Comment | Organism |
---|---|---|
SHC | - |
Bradyrhizobium japonicum |
SHC | - |
Rhodopseudomonas palustris |
SHC | - |
Streptomyces peucetius |
SHC | - |
Zymomonas mobilis |
SHC | - |
Methylococcus capsulatus |
SHC | - |
Tetrahymena thermophila |
SHC | - |
Alicyclobacillus acidocaldarius |
squalene-hopene cyclase | - |
Bradyrhizobium japonicum |
squalene-hopene cyclase | - |
Alicyclobacillus acidocaldarius |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
28 | - |
- |
Bradyrhizobium japonicum |
30 | - |
- |
Rhodopseudomonas palustris |
30 | - |
- |
Zymomonas mobilis |
30 | - |
- |
Tetrahymena thermophila |
35 | - |
- |
Streptomyces peucetius |
40 | - |
- |
Methylococcus capsulatus |
60 | - |
- |
Alicyclobacillus acidocaldarius |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
- |
Zymomonas mobilis |
6 | - |
- |
Alicyclobacillus acidocaldarius |
6.5 | - |
- |
Bradyrhizobium japonicum |
6.5 | - |
- |
Rhodopseudomonas palustris |
6.8 | - |
- |
Streptomyces peucetius |
6.8 | - |
- |
Methylococcus capsulatus |
7 | - |
- |
Tetrahymena thermophila |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 8 | activity range | Rhodopseudomonas palustris |
General Information | Comment | Organism |
---|---|---|
evolution | structure-function relationships of squalene-hopene cyclases, the DXDD motif, which is typical for all squalene-hopene cyclases, overview | Bradyrhizobium japonicum |
evolution | structure-function relationships of squalene-hopene cyclases, the DXDD motif, which is typical for all squalene-hopene cyclases, overview | Rhodopseudomonas palustris |
evolution | structure-function relationships of squalene-hopene cyclases, the DXDD motif, which is typical for all squalene-hopene cyclases, overview | Streptomyces peucetius |
evolution | structure-function relationships of squalene-hopene cyclases, the DXDD motif, which is typical for all squalene-hopene cyclases, overview | Zymomonas mobilis |
evolution | structure-function relationships of squalene-hopene cyclases, the DXDD motif, which is typical for all squalene-hopene cyclases, overview | Methylococcus capsulatus |
evolution | structure-function relationships of squalene-hopene cyclases, the DXDD motif, which is typical for all squalene-hopene cyclases, overview | Tetrahymena thermophila |
evolution | structure-function relationships of squalene-hopene cyclases, the DXDD motif, which is typical for all squalene-hopene cyclases, overview | Alicyclobacillus acidocaldarius |
evolution | enzyme distribution in the different taxa, overview | Bradyrhizobium japonicum |
evolution | enzyme distribution in the different taxa, overview | Alicyclobacillus acidocaldarius |
metabolism | the enzyme is involved in biosynthesis of hopanoids, members of a large group of cyclic triterpenoic compounds that have important functions in many prokaryotic and eukaryotic organisms | Bradyrhizobium japonicum |
metabolism | the enzyme is involved in biosynthesis of hopanoids, members of a large group of cyclic triterpenoic compounds that have important functions in many prokaryotic and eukaryotic organisms | Rhodopseudomonas palustris |
metabolism | the enzyme is involved in biosynthesis of hopanoids, members of a large group of cyclic triterpenoic compounds that have important functions in many prokaryotic and eukaryotic organisms | Streptomyces peucetius |
metabolism | the enzyme is involved in biosynthesis of hopanoids, members of a large group of cyclic triterpenoic compounds that have important functions in many prokaryotic and eukaryotic organisms | Zymomonas mobilis |
metabolism | the enzyme is involved in biosynthesis of hopanoids, members of a large group of cyclic triterpenoic compounds that have important functions in many prokaryotic and eukaryotic organisms | Methylococcus capsulatus |
metabolism | the enzyme is involved in biosynthesis of hopanoids, members of a large group of cyclic triterpenoic compounds that have important functions in many prokaryotic and eukaryotic organisms | Alicyclobacillus acidocaldarius |
metabolism | the nezym eis involved in biosynthesis of hopanoids, members of a large group of cyclic triterpenoic compounds that have important functions in many prokaryotic and eukaryotic organisms | Tetrahymena thermophila |
metabolism | the enzyme converts squalene to hopanol, EC 4.2.1.129, and to hopene, EC 5.4.99.17, but not to tetrahymanol, EC 4.2.1.123, pathway overview | Alicyclobacillus acidocaldarius |
metabolism | the enzyme converts squalene to tetrahymanol, EC 4.2.1.123, to hopene, EC 5.4.99.17, and to hopanol, EC 4.2.1.129, pathway overview | Bradyrhizobium japonicum |
additional information | structure-function relationships of squalene-hopene cyclases, overview | Bradyrhizobium japonicum |
additional information | structure-function relationships of squalene-hopene cyclases, overview | Rhodopseudomonas palustris |
additional information | structure-function relationships of squalene-hopene cyclases, overview | Streptomyces peucetius |
additional information | structure-function relationships of squalene-hopene cyclases, overview | Zymomonas mobilis |
additional information | structure-function relationships of squalene-hopene cyclases, overview | Methylococcus capsulatus |
additional information | structure-function relationships of squalene-hopene cyclases, overview | Tetrahymena thermophila |
additional information | structure-function relationships of squalene-hopene cyclases, overview. A large central cavity represents the catalytic site in Alicyclobacillus acidocaldarius enzyme that takes up and orientates the squalene molecule. The channel and active-site cavity inside the protein are separated by a narrow constriction buildup of four amino acids, D376, F166, C435, and F434, that appear to block access to the active site | Alicyclobacillus acidocaldarius |
additional information | structure-function relationships of SHCs, active site structure, overview | Bradyrhizobium japonicum |
additional information | structure-function relationships of SHCs, active site structure, overview. A protruding part in the center of the nonpolar region contains a lipophilic channel and directs the substrate to the active-site cavity inside the protein. The channel and cavity are separated by a narrow constriction buildup of four amino acids, D376, F166, C435, and F434, that appear to block access to the active site. Residues C435 and F434 are part of a loop that seems to be flexible enough to permit passage of the substrate and the product | Alicyclobacillus acidocaldarius |