Metabolism of synthetic inositol trisphosphate analogs.
A series of synthetic analogs was employed to explore structure-activity relationships in the metabolism of the second messenger inositol trisphosphate (IP3) in vascular tissue. Cytosolic IP3-5-phosphatase activity was purified approximately 240-fold from bovine aorta. All synthetic analogs tested were apparent competitive inhibitors of the 5-phosphatase activity. The order of potency was DL-1,3,4,5-IP3 greater than D-1,4,5-IP3 greater than DL-1,3,4-IP3 greater than L-1,4,5-IP3 greater than 1,3,5-IP3 greater than DL-6-methoxy-1,4,5-IP3 greater than DL-2,4,5-IP3 greater than DL-1,2,4-cyclohexane-P3. The least potent analogs had Ki values only 11 times higher than the apparent Km of the substrate D-1,4,5-[3H]IP3. However, only three synthetic compounds, DL-1,3,4,5-IP4, D-1,4,5-IP3, and DL-2,4,5-IP3, could serve as substrates for the 5-phosphatase. IP3 kinase activity in the same tissue exhibited considerably more selectivity with respect to inhibition by IP3 analogs. D-1,4,5-IP3 was about 30 times more potent than DL-1,3,4,5-IP4 and 100-1000 times more potent than the other compounds tested. The function of the IP3 receptor was evaluated by measuring labeled calcium mobilization in permeabilized bovine aortic smooth muscle cells in culture. While all analogs tested were full agonists, vast differences in potency were observed. D-1,4,5-IP3 was about 30 times more potent than DL-2,4,5-IP3 and 100-2000 times more potent than the other analogs tested. The results suggest that IP3-5-phosphatase activity is relatively nonselective in the binding of inositol polyphosphates, while IP3 kinase activity and the IP3 receptor exhibit great selectivity in the recognition of these compounds.
Polokoff
M A
MA
Department of Biological Chemistry, Merck Sharp and Dohme Research Laboratories, West Point, Pennsylvania 19486.
Bencen
G H
GH
Vacca
J P
JP
deSolms
S J
SJ
Young
S D
SD
Huff
J R
JR
D000818:Animals; D001011:Aorta / Q000201:enzymology; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels; D002417:Cattle; D002478:Cells, Cultured; D003600:Cytosol / Q000201:enzymology; D006868:Hydrolysis; D015544:Inositol 1,4,5-Trisphosphate; D053496:Inositol 1,4,5-Trisphosphate Receptors; D007295:Inositol Phosphates / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D000072181:Inositol Polyphosphate 5-Phosphatases; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism; D010770:Phosphotransferases / Q000037:antagonists & inhibitors / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor); D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D013403:Sugar Phosphates / Q000378:metabolism*
D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D007295:Inositol Phosphates; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; D013403:Sugar Phosphates; D015544:Inositol 1,4,5-Trisphosphate; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C051038:Inositol 1,4,5-trisphosphate 3-kinase; D010744:Phosphoric Monoester Hydrolases; D000072181:Inositol Polyphosphate 5-Phosphatases; D002118:Calcium