Synthesis and biological properties of 2-substituted myo-inositol 1,4,5-trisphosphate analogues directed toward affinity chromatography and photoaffinity labeling.
A series of myo-inositol 1,4,5-trisphosphate analogues with the 2-acyl substituents p-aminobenzoyl (7), p-azidobenzoyl (8), 4-(5-[2-(benzamido)ethyl]-2-hydroxyphenylazo)benzoyl (9), and cis,trans-4-aminocyclohexylcarbonyl (10) were synthesised and examined for their effects on the 5-phosphatase, the 3-kinase, the tritiated trisphosphate-binding activity, and the Ca(2+)-releasing activity. Each analogue inhibited the hydrolysis of D-[5-32P]Ins(1,4,5)P3 and the phosphorylation of D-[3H]Ins(1,4,5)P3, catalysed by erythrocyte ghosts and brain cytosol, respectively. The analogues acted as full agonists in releasing Ca2+ from permeabilised cells and also inhibited the binding of D-[3H]Ins(1,4,5)P3 to cerebellum microsomes. The analogues 7 and 10 were utilised for immobilisation of the trisphosphate on Sepharose and the subsequent affinity chromatography effected purification of the above proteins. A photoaffinity probe, the appendage of which acted as the photoaffinity probe as well as a non-radioactive molecular marker, was also derived from the analogue 7.
Ozaki
S
S
Department of Applied Chemistry, Faculty of Engineering, Ehime University, Matsuyama, Japan.
Watanabe
Y
Y
Ogasawara
T
T
Hirata
M
M
Kanematsu
T
T
D000818:Animals; D001921:Brain / Q000187:drug effects / Q000201:enzymology; D002118:Calcium / Q000378:metabolism; D002384:Catalysis; D002846:Chromatography, Affinity; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology; D004910:Erythrocyte Membrane / Q000187:drug effects; D006801:Humans; D006868:Hydrolysis; D015544:Inositol 1,4,5-Trisphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D015394:Molecular Structure; D010766:Phosphorylation; D051381:Rats
D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium