Ca2+ dependent activation of rat uterine glycerylphosphorylcholine diesterase: presence of a positive modulator protein in uterine secretion.
The rat uterine secretory enzyme glycerylphosphorylcholine (GPC) diesterase (EC 3.1.4.2) had been purified and characterized previously with respect to its mol. wt., size, amino acid, carbohydrate composition and estrogen inducible properties. This enzyme is observed to have exclusive specificity for GPC and exhibits characteristic hyperbolic kinetics with Ca2+ in an ethyleneglycolbis N'N'N'N' tetraacetic acid (EGTA) buffered system. Ca2+ reduces Km of the enzyme for GPC from 0.65 to 0.25 mM. The Km for GPC of the partially purified enzyme is found to be 0.35 mM without addition of calcium which indicates the presence of a positive modulator of the enzyme in this fraction. Based on this rationale, a protein activating factor for the enzyme was isolated from this fraction which has a native size of 18 KD as observed on Sephacryl S-200 chromatography and strikingly stimulates enzyme activity at around 0.55 microM.
Mitra
J
J
Indian Institute of Chemical Biology, Calcutta.
Chowdhury
M
M
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D004492:Edetic Acid / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004789:Enzyme Activation; D005260:Female; D005997:Glycerylphosphorylcholine / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D008970:Molecular Weight; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D011359:Proestrus; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D013379:Substrate Specificity; D014599:Uterus / Q000737:chemistry / Q000201:enzymology*
D002147:Calmodulin; D011506:Proteins; D004533:Egtazic Acid; D005997:Glycerylphosphorylcholine; D004492:Edetic Acid; D010727:Phosphoric Diester Hydrolases; C020677:glycerophosphocholine phosphodiesterase; D008274:Magnesium; D002118:Calcium