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Literature summary for 2.7.1.127 extracted from

  • Woodring, P.J.; Garrison, J.C.
    Expression, purification, and regulation of two isoforms of the inositol 1,4,5-trisphosphate 3-kinase (1997), J. Biol. Chem., 272, 30447-30454.
    View publication on PubMed
Search for more literature for 2.7.1.127

Activating Compound

Activating Compound Comment Organism Structure
protein kinase A isoforms A and B are differentially regulated via phosphorylation by the cAMP-dependent protein kinase, isoform A: phosphorylated to the extent of 0.9 mol/mol, 2.5fold stimulation in the absence of Ca2+/calmodulin, 1.5fold stimulation in the presence of Ca2+/calmodulin, isoform B: phosphorylated to the extent of 1 mol/mol, no effect on activity in the absence of Ca2+/calmodulin, 45% inhibition in the presence of Ca2+/calmodulin Rattus norvegicus

Cloned(Commentary)

Cloned (Comment) Organism
cDNAs encoding enzyme isoforms A from brain and B are cloned and overexpressed in B31 cells, a rat-1 fibroblast cell line expressing pp60v-src Rattus norvegicus

General Stability

General Stability Organism
2-mercaptoethanol and Triton X-100 or CHAPS stabilize during purification Rattus norvegicus
bovine serum albumin stabilizes dilute enzyme solutions Rattus norvegicus
isoform A is more susceptible to proteolysis during purification than isoform B Rattus norvegicus

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ isoforms A and B, above 0.01 mM Rattus norvegicus
protein kinase A isoforms A and B are differentially regulated via phosphorylation by the cAMP-dependent protein kinase, isoform A: stimulation in the presence or absence of Ca2+/calmodulin, isoform B: no effect on activity in the absence of Ca2+/calmodulin, 45% inhibition in the presence of Ca2+/calmodulin Rattus norvegicus
Protein kinase C isoforms A and B are differentially regulated via phosphorylation by protein kinase C, isoform A: phosphorylated to the extent of 2 mol/mol, 72% inhibition in the absence of Ca2+/calmodulin, isoform B: phosphorylated to the extent of 2.7 mol/mol, no effect on activity in the absence of Ca2+/calmodulin, both isoforms are inhibited by 70% in the presence of Ca2+/calmodulin Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.002 0.005 1D-myo-inositol 1,4,5-trisphosphate pH 7.5, 37°C, recombinant isoenzymes A and B Rattus norvegicus

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
 Rattus norvegicus 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ isoforms A and B are differentially regulated by Ca2+/calmodulin, isoform A: 2.5fold stimulation, isoform B: 20fold stimulation Rattus norvegicus
Ca2+ activation by Ca2+/calmodulin Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
59000
-
 x * 59000, recombinant isoform A, x * 92000, recombinant isoform B, SDS-PAGE Rattus norvegicus
92000
-
 x * 59000, recombinant isoform A, x * 92000, recombinant isoform B, SDS-PAGE Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + 1D-myo-inositol 1,4,5-trisphosphate Rattus norvegicus differential expression and regulation of the two enzyme isoforms A and B provides multiple mechanisms for regulating the cytosolic level of inositol 1,4,5-trisphosphate in cells, regulation mechanisms ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate
-
 ?
ATP + 1D-myo-inositol 1,4,5-trisphosphate Rattus norvegicus inositol 1,4,5-trisphosphate is a second messenger ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate
-
 ?

Organic Solvent Stability

Organic Solvent Comment Organism
Triton X-100 stabilizes Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
 2 isoenzymes: A and B
-

Posttranslational Modification

Posttranslational Modification Comment Organism
phosphoprotein isoforms A and B are differentially regulated via phosphorylation by protein kinase C and the cAMP-dependent protein kinase, potential phosphorylation sites Rattus norvegicus

Purification (Commentary)

Purification (Comment) Organism
recombinant isoforms A and B, overexpressed in B31 rat fibroblast, isoform A: 11005fold, isoform B: 43806fold Rattus norvegicus

Source Tissue

Source Tissue Comment Organism Textmining
brain isoform A Rattus norvegicus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
15.77
-
 pH 7.5, 37°C, recombinant isoform B Rattus norvegicus
36.32
-
 pH 7.5, 37°C, recombinant isoform A Rattus norvegicus

Storage Stability

Storage Stability Organism
-70°C, buffer containing detergent and reducing agent, pH 7-8, at least 3 months, stable Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 1D-myo-inositol 1,4,5-trisphosphate multiple regulation mechanisms of enzyme isoform A and B Rattus norvegicus ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate
-
 ?
ATP + 1D-myo-inositol 1,4,5-trisphosphate differential expression and regulation of the two enzyme isoforms A and B provides multiple mechanisms for regulating the cytosolic level of inositol 1,4,5-trisphosphate in cells, regulation mechanisms Rattus norvegicus ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate
-
 ?
ATP + 1D-myo-inositol 1,4,5-trisphosphate inositol 1,4,5-trisphosphate is a second messenger Rattus norvegicus ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate
-
 ?

Subunits

Subunits Comment Organism
? x * 59000, recombinant isoform A, x * 92000, recombinant isoform B, SDS-PAGE Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Rattus norvegicus
Calmodulin activation by Ca2+/calmodulin Rattus norvegicus
Calmodulin isoforms A and B are differentially regulated by Ca2+/calmodulin, isoform A: 2.5fold stimulation, isoform B: 20fold stimulation Rattus norvegicus