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Literature summary for 4.6.1.13 extracted from

  • Kravchuk, A.V.; Zhao, L.; Bruzik, K.S.; Tsai, M.D.
    Engineering a catalytic metal binding site into a calcium-independent phosphatidylinositol-specific phospholipase C leads to enhanced stereoselectivity (2003), Biochemistry, 42, 2422-2430.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
D33N/R69D PI-PLC double mutant, 50fold activation by 1 mM Ca2+ Bacillus sp. (in: Bacteria)
R69A mutation of the catalytic Arg-69, not activated by Ca2+ Bacillus sp. (in: Bacteria)
R69C mutation of the catalytic Arg-69, not activated by Ca2+ Bacillus sp. (in: Bacteria)
R69D reduced activity compared with wild-type enzyme, mutant is activated by Ca2+, mutation engineers a catalytic metal binding site into the calcium-independent PI-PLC leading to enhanced stereoselectivity Bacillus sp. (in: Bacteria)
R69E mutation of the catalytic Arg-69, inactive mutant, not activated by Ca2+ Bacillus sp. (in: Bacteria)
R69G mutation of the catalytic Arg-69, not activated by Ca2+ Bacillus sp. (in: Bacteria)
R69N mutation of the catalytic Arg-69, not activated by Ca2+ Bacillus sp. (in: Bacteria)

Inhibitors

Inhibitors Comment Organism Structure
Cd2+ severe inhibition of wild-type PI-PLC Bacillus sp. (in: Bacteria)
Co2+ severe inhibition of wild-type PI-PLC Bacillus sp. (in: Bacteria)
Ni2+ severe inhibition of wild-type PI-PLC Bacillus sp. (in: Bacteria)
Zn2+ severe inhibition of wild-type PI-PLC, complete inhibition of R69D mutant below 1 mM Bacillus sp. (in: Bacteria)

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ 41fold activation of R69D mutant, slight activation of wild-type PI-PLC Bacillus sp. (in: Bacteria)
Cd2+ activates R69D mutant at low concentrations, no activation of wild-type PI-PLC Bacillus sp. (in: Bacteria)
Co2+ activates R69D mutant at low concentrations, no activation of wild-type PI-PLC Bacillus sp. (in: Bacteria)
Li+ 5fold activation of R69D mutant, slight activation of wild-type PI-PLC Bacillus sp. (in: Bacteria)
Mg2+ 35fold activation of R69D mutant, slight activation of wild-type PI-PLC Bacillus sp. (in: Bacteria)
Mn2+ activates R69D mutant at low concentrations, slight activation of wild-type PI-PLC Bacillus sp. (in: Bacteria)
additional information wild-type PI-PLC is calcium-independent Bacillus sp. (in: Bacteria)
Sr2+ 9fold activation of R69D mutant, slight activation of wild-type PI-PLC Bacillus sp. (in: Bacteria)

Organism

Organism UniProt Comment Textmining
Bacillus sp. (in: Bacteria)
-
-
-

Purification (Commentary)

Purification (Comment) Organism
PI-PLC mutants Bacillus sp. (in: Bacteria)

Reaction

Reaction Comment Organism Reaction ID
1-phosphatidyl-1D-myo-inositol = 1D-myo-inositol 1,2-cyclic phosphate + 1,2-diacyl-sn-glycerol mechanism Bacillus sp. (in: Bacteria)

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
values for wild-type and several mutant PI-PLCs in presence of 0.1 mM or 1 mM Ca2+ and in absence of Ca2+ Bacillus sp. (in: Bacteria)
3560
-
pH 7.5, 37°C, wild-type PI-PLC, in absence of Ca2+ Bacillus sp. (in: Bacteria)

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-phosphatidyl-1D-myo-inositol PLC accepts only nonphosphorylated phosphatidylinositol substrates and produces cyclic inositol phosphate as final product, which is hydrolyzed at a 1000fold lower rate, catalytic mechanism, uses a guanidinium group of Arg-69 during catalysis Bacillus sp. (in: Bacteria) 1D-myo-inositol 1,2-cyclic phosphate + diacylglycerol
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Bacillus sp. (in: Bacteria)

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Bacillus sp. (in: Bacteria)