Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in COS-7 cells or HEK-293 cells. The wild-type and S406A and S406T mutants are expressed on the plasma membrane in glycosylphosphatidylinositol-anchored form, the S406P mutant is notz and is retained in a perinuclear location. Expression in baculovirus infected cells in a glycophosphatidyl-anchored form, whereas a truncated form, lacking the putative signal sequence for glycosylphosphatidyl anchoring is secreted at high levels into the medium. Both forms have lower molecular masses than native placental enzyme indicating a minimal glycosylation | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
E260A | mutation reduces the ratio of turnover-numver to Km-value by 104fold and further decreases stability.Addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wild-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297% | Homo sapiens |
E260Q | mutation has minimal effects on kinetic parameters, but decreased heat stability | Homo sapiens |
E264Q | mutation results in a 10000fold decrease in activity, but the enzyme still binds to p-aminobenzoylarginine-Sepharose and is resistant to trypsin treatment, indicating that the protein is folded properly | Homo sapiens |
S406A | very similar to the wild-type enzyme with regard to expression and release by phosphatidylinositol-specific phospholipase C | Homo sapiens |
S406P | the wild-type and S406A and S406T mutants are expressed on the plasma membrane in glycosylphosphatidylinositol-anchored form, the S406P mutant is not and is retained in a perinuclear location | Homo sapiens |
S406T | very similar to the wild-type enzyme with regard to expression and release by phosphatidylinositol-specific phospholipase C | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,10-phenanthroline | 1 mM, 96% inhibition | Homo sapiens | |
Guanidinoethyl mercaptosuccinic acid | 1 mM, 97% inhibition | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.059 | - |
Dansyl-Ala-Arg | wild-type enzyme lacking the C-terminal hydrophobic membrane anchor signal | Homo sapiens | |
0.102 | - |
Dansyl-Ala-Arg | mutant enzyme E260Q | Homo sapiens | |
0.28 | - |
Dansyl-Ala-Arg | mutant enzyme E260A | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
plasma membrane | expression of wild-type and mutant enzymes in COS-7 cells or HEK-293 cells.The wild-type and S406A and S406T mutants are expressed on the plasma membrane in glycosylphosphatidylinositol-anchored form, the S406P mutant is not and is retained in a perinuclear location | Homo sapiens | 5886 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
KCl | addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297% | Homo sapiens | |
KNO3 | addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297% | Homo sapiens | |
Na2SO4 | addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297% | Homo sapiens | |
NaCl | addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297% | Homo sapiens | |
NaNO3 | addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297% | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
50000 | - |
x * 50000, SDS-PAGE | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | the wild-type enzyme lacking the C-terminal hydrophobic membrane anchor signal contains only 5.4% carbohydrate | Homo sapiens |
glycoprotein | a truncated form, lacking the putative signal sequence for glycosylphosphatidyl anchoring is secreted at high levels into the medium | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
placenta | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Dansyl-Ala-Arg + H2O | - |
Homo sapiens | Dansyl-Ala + Arg | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 50000, SDS-PAGE | Homo sapiens |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
wild-type enzyme and mutant enzymes E260Q and E260A are stable | Homo sapiens |
45 | - |
15 min, 50% inactivation of mutant enzyme E260A | Homo sapiens |
49 | - |
15 min, 15 min, 50% inactivation of mutant enzyme E260Q | Homo sapiens |
50 | - |
15 min, complete inactivation of mutant enzyme E260A | Homo sapiens |
53 | - |
15 min, wild-type enzyme retains 50% of its activity | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.233 | - |
Dansyl-Ala-Arg | mutant enzyme E260A | Homo sapiens | |
5.12 | - |
Dansyl-Ala-Arg | wild-type enzyme lacking the C-terminal hydrophobic membrane anchor signal | Homo sapiens | |
6.02 | - |
Dansyl-Ala-Arg | mutant enzyme E260Q | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | 7.5 | - |
Homo sapiens |