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Literature summary for 2.7.8.11 extracted from
- The active site of yeast phosphatidylinositol synthase Pis1 is facing the cytosol (2015), Biochim. Biophys. Acta, 1851, 629-640.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene PIS1, recombinant expression of wild-type and mutant V5-His6-tagged enzyme Pis1 in a diploid pis1DELTA::kanMX/PIS1 strain | Saccharomyces cerevisiae |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K65C | site-directed mutagenesis, cysteine installation for substituted cysteine analysis method, SCAM | Saccharomyces cerevisiae |
| additional information | an addition of a KXKXX motif reduces the activity of Pis1 by a factor of 2 | Saccharomyces cerevisiae |
| N67C | site-directed mutagenesis, cysteine installation for substituted cysteine analysis method, SCAM | Saccharomyces cerevisiae |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| dodecylmaltoside | DDM, enzyme Pis1 is highly sensitive to non-denaturing detergent, and low concentrations (0.05%) of dodecylmaltoside change the accessibility of single substituted Cys in the mutated active site of an otherwise cysteine free version of enzyme Pis1. Slightly higher detergent concentrations inactivate the enzyme. Complete inactivation of Pis1 by 1% DDM is the consequence of a rather drastic loss of 3D structure | Saccharomyces cerevisiae |  |
| additional information | the Pis1 enzyme structure is strongly affected by mild detergents. Cys53 and Cys154 are residing close to the bottom of the cytosolic cavity of Pis1 and are accessible for small detergent molecules | Saccharomyces cerevisiae |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| endoplasmic reticulum membrane | membrane topology of enzyme Pis, overview. The enzyme central sequence contains 6 transmembrane helices | Saccharomyces cerevisiae | 5789 | - |
| microsome | - |
Saccharomyces cerevisiae | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Saccharomyces cerevisiae | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| CDP-diacylglycerol + myo-inositol | Saccharomyces cerevisiae | - |
CMP + 1-phosphatidyl-1D-myo-inositol | - |
? | |
| CDP-diacylglycerol + myo-inositol | Saccharomyces cerevisiae BY4742 | - |
CMP + 1-phosphatidyl-1D-myo-inositol | - |
? | |
| additional information | Saccharomyces cerevisiae | cytosolically made inositol and CDP-diacylglycerol can access the active site of the yeast PI synthase Pis1 from the cytosolic side | ? | - |
? | |
| additional information | Saccharomyces cerevisiae BY4742 | cytosolically made inositol and CDP-diacylglycerol can access the active site of the yeast PI synthase Pis1 from the cytosolic side | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Saccharomyces cerevisiae | P06197 | - |
- |
| Saccharomyces cerevisiae BY4742 | P06197 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutant V5-His6-tagged enzyme Pis1 partially by microsome preparation | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| CDP-diacylglycerol + myo-inositol | - |
Saccharomyces cerevisiae | CMP + 1-phosphatidyl-1D-myo-inositol | - |
? | |
| CDP-diacylglycerol + myo-inositol | - |
Saccharomyces cerevisiae BY4742 | CMP + 1-phosphatidyl-1D-myo-inositol | - |
? | |
| additional information | cytosolically made inositol and CDP-diacylglycerol can access the active site of the yeast PI synthase Pis1 from the cytosolic side | Saccharomyces cerevisiae | ? | - |
? | |
| additional information | cytosolically made inositol and CDP-diacylglycerol can access the active site of the yeast PI synthase Pis1 from the cytosolic side | Saccharomyces cerevisiae BY4742 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| phosphatidylinositol synthase | - |
Saccharomyces cerevisiae |
| PI synthase | - |
Saccharomyces cerevisiae |
| PIS1 | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 22 | - |
assay at rrom temperature | Saccharomyces cerevisiae |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.2 | - |
assay at | Saccharomyces cerevisiae |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the CDP-alcohol phosphatidyltransferase (CAPT) superfamily. Members of the family share the so-called CAPT motif, which forms the active site of all these enzymes. The CAPT motif of phosphatidylinositol synthase Pis1 is located to the cytosolic side, structure comparisons, overview | Saccharomyces cerevisiae |
| malfunction | removal of the endoplasmic reticulum retrieval sequence from wild-type enzyme Pis1 enhances its activity, suggesting an influence of the lipid environment | Saccharomyces cerevisiae |
| additional information | the Pis1 enzyme structure is strongly affected by mild detergents. Modeling of enzyme Pis1 structure, overview | Saccharomyces cerevisiae |
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