TY - JOUR
T1 - Disaccharide polyphosphates based upon adenophostin A activate hepatic D-myo-inositol 1,4,5-trisphosphate receptors
AU - Marchant, Jonathan S.
AU - Beecroft, Mike D.
AU - Riley, Andrew M.
AU - Jenkins, David J.
AU - Marwood, Rachel D.
AU - Taylor, Colin W.
AU - Potter, Barry V.L.
PY - 1997/10/21
Y1 - 1997/10/21
N2 - The glyconucleotides adenophostin A and B are the most potent known agonists at type 1 inositol trisphosphate [Ins(1,4,5)P3] receptors, although their stuctures differ markedly from that of Ins(1,4,5)P3. Equilibrium competition binding with [3H]Ins(1,4,5)P3 and unidirectional 45Ca2+ flux measurements were used to examine the effects of adenophostin A in hepatocytes, which express predominantly type 2 Ins(1,4,5)P3 receptors. Both Ins(1,4,5)P3 (K(d) = 8.65 ± 0.98 nM) and adenophostin A (K(d) = 0.87 ± 0.20 nM) bound to a single class of [3H]Ins(1,4,5)P3-binding site and each fully mobilized the same intracellular Ca2+ pool; although, adenophostin A (EC50 = 10.9 ± 0.7 nM) was more potent than Ins(1,4,5)P3 (EC50 = 153 ±11 nM). Working on the assumption that it is the phosphorylated glucose component of the adenophostins that mimics the critical features of Ins(1,4,5)P3, we synthesized various phosphorylated disaccharide analogs containing this structure. The novel disaccharide-based analogs, sucrose 3,4,3'-trisphosphate [Sucr(3,4,3')P3], α,α'-trehalose 3,4,3',4'- tetrakisphosphate [Trehal-(3,4,3',4')P4], α,α'-trehalose 2,4,3',4'- tetrakisphosphate [Trehal(2,4,3',4')P4], and methyl 3-O-(α-D- glucopyranosyl)-β-D-ribofuranoside 2,3',4'-trisphosphate [Rib(2,3',4')P3], were all able to mobilize the same intracellular Ca2+ pool as Ins(1,4,5)P3 and adenophostin A; although, none was as potent as adenophostin A. The rank order of potency of the analogs, adenophostin A > Ins(1,4,5)P3 ≃ Rib(2,3',4')-P3 > Trehal(2,4,3',4')P4 > Glc(2'3,4)P3 ≃ Trehal(3,4,3',4')P4 > Sucr(3,4,3')P3, was the same in radioligand binding and functional assays of hepatic Ins(1,4,5)P3 receptors. Both Rib(2,3',4')P3, which was as potent as Ins(1,4,5)P3, and Trehal(2,4,3',4')P4 bound with significantly higher affinity (~27 and ~3- fold, respectively) than the only active carbohydrate agonist of Ins(1,4,5)P3 receptors previously examined [Glc(2',3,4)P3]. We conclude that phosphorylated disaccharides provide novel means of developing high- affinity ligands of Ins(1,4,5)P3 receptors.
AB - The glyconucleotides adenophostin A and B are the most potent known agonists at type 1 inositol trisphosphate [Ins(1,4,5)P3] receptors, although their stuctures differ markedly from that of Ins(1,4,5)P3. Equilibrium competition binding with [3H]Ins(1,4,5)P3 and unidirectional 45Ca2+ flux measurements were used to examine the effects of adenophostin A in hepatocytes, which express predominantly type 2 Ins(1,4,5)P3 receptors. Both Ins(1,4,5)P3 (K(d) = 8.65 ± 0.98 nM) and adenophostin A (K(d) = 0.87 ± 0.20 nM) bound to a single class of [3H]Ins(1,4,5)P3-binding site and each fully mobilized the same intracellular Ca2+ pool; although, adenophostin A (EC50 = 10.9 ± 0.7 nM) was more potent than Ins(1,4,5)P3 (EC50 = 153 ±11 nM). Working on the assumption that it is the phosphorylated glucose component of the adenophostins that mimics the critical features of Ins(1,4,5)P3, we synthesized various phosphorylated disaccharide analogs containing this structure. The novel disaccharide-based analogs, sucrose 3,4,3'-trisphosphate [Sucr(3,4,3')P3], α,α'-trehalose 3,4,3',4'- tetrakisphosphate [Trehal-(3,4,3',4')P4], α,α'-trehalose 2,4,3',4'- tetrakisphosphate [Trehal(2,4,3',4')P4], and methyl 3-O-(α-D- glucopyranosyl)-β-D-ribofuranoside 2,3',4'-trisphosphate [Rib(2,3',4')P3], were all able to mobilize the same intracellular Ca2+ pool as Ins(1,4,5)P3 and adenophostin A; although, none was as potent as adenophostin A. The rank order of potency of the analogs, adenophostin A > Ins(1,4,5)P3 ≃ Rib(2,3',4')-P3 > Trehal(2,4,3',4')P4 > Glc(2'3,4)P3 ≃ Trehal(3,4,3',4')P4 > Sucr(3,4,3')P3, was the same in radioligand binding and functional assays of hepatic Ins(1,4,5)P3 receptors. Both Rib(2,3',4')P3, which was as potent as Ins(1,4,5)P3, and Trehal(2,4,3',4')P4 bound with significantly higher affinity (~27 and ~3- fold, respectively) than the only active carbohydrate agonist of Ins(1,4,5)P3 receptors previously examined [Glc(2',3,4)P3]. We conclude that phosphorylated disaccharides provide novel means of developing high- affinity ligands of Ins(1,4,5)P3 receptors.
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U2 - 10.1021/bi971397v
DO - 10.1021/bi971397v
M3 - Article
C2 - 9335535
AN - SCOPUS:0030722963
SN - 0006-2960
VL - 36
SP - 12780
EP - 12790
JO - Biochemistry
JF - Biochemistry
IS - 42
ER -