Stereospecific recognition of inositol 1,4,5-trisphosphate analogs by the phosphatase, kinase, and binding proteins.
A series of DL-inositol 1,4,5-trisphosphate (IP3) analogs, with a bulky substitutent on the 2nd carbon of the inositol ring, has been synthesized. These compounds exert biological activities with only minor reduction in potency, in several assay systems (Hirata, M., Watanabe, Y., Ishimatsu, T., Ikebe, T., Kimura, Y., Yamaguchi, K., Ozaki, S., and Koga, T. (1989) J. Biol. Chem. 264, 20303-20308). Two analogs with aminocyclohexanecarbonyl (designated as analog 206) or aminobenzoyl group (analog 209) were separated into individual optical isomers and examined for stereospecificity in recognition by IP3-5-phosphatase, IP3-3-kinase and IP3 binding activity. IP3-5-phosphatase activity of erythrocyte ghosts was competitively inhibited by L-209 with a lower Ki value than D-IP3, but with a higher Ki value by L-206. D-Isomers of both analogs at 100 microM failed to inhibit the hydrolysis of D-[3H]IP3. On the other hand, D-isomers but not L-isomers of both analogs were as potent as D-IP3 in the recognition by IP3-3-kinase of rat brain cytosol and only the D-isomer of analog 206 could serve as substrate for the kinase. Also D-isomers of both analogs were equipotent to D-IP3 in displacing [3H]IP3 binding to rat cerebellum microsomes. These observations suggest that the IP3 analogs we synthesized are stereospecifically recognized by three IP3-recognizable proteins, but the phosphatase recognizes opposite isomers. Such being the case, the second hydroxyl group of D-IP3 may be involved in the recognition by IP3-5-phosphatase, but not by IP3-3-kinase and binding sites.
Hirata
M
M
Department of Biochemistry, Faculty of Dentistry, Kyushu University, Fukuoka, Japan.
Yanaga
F
F
Koga
T
T
Ogasawara
T
T
Watanabe
Y
Y
Ozaki
S
S
D000818:Animals; D001921:Brain / Q000201:enzymology; D015220:Calcium Channels; D002531:Cerebellum / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D004910:Erythrocyte Membrane / Q000201:enzymology; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D053496:Inositol 1,4,5-Trisphosphate Receptors; D000072181:Inositol Polyphosphate 5-Phosphatases; D007700:Kinetics; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010770:Phosphotransferases / Q000037:antagonists & inhibitors / Q000097:blood*; D017853:Phosphotransferases (Alcohol Group Acceptor); D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear; D013379:Substrate Specificity
D015220:Calcium Channels; C506926:ITPR1 protein, human; D053496:Inositol 1,4,5-Trisphosphate Receptors; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; 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