[Use of spectral characteristics of ryodipine in the study of dihydropyridine receptor complex of neuronal membranes].
A fluorescent dihydropyridine (DHP) derivative, ryodipine, was used to study structural characteristics of the DHP-sensitive Ca-channels in nerve terminals (synaptosomes) isolated from the rat cerebral cortex. It was found that an inductive resonance energy transfer from membrane proteins to ryodipine occurred in synaptosomal membranes. Two groups of membrane proteins differentially accessible to ryodipine were found by quenching of their own fluorescence. The percentage of group I proteins (20%) whose fluorescence was quenched by up to 1 microM ryodipine, was increased by 50% upon K(+)-depolarization and remained unchanged upon the addition of 100 microM Ni2+, whereas the addition of 100 microM Cd2+ prevented the increase induced by K(+)-depolarization. Nifedipine and nicardipine competed with ryodipine for the DHP receptor as evidenced by the change in percentage of group I proteins. The percentage of group II proteins (50% at 10 microM ryodipine) remained unchanged during various functional alterations of the synaptosomal membranes. Model experiments on proteoliposomes demonstrated that binding of ryodipine to synaptosomal membranes was due mainly to the hydrophobicity of DHP but not the ligand-receptor interaction. Nonetheless we that the membrane proteins-ryodipine system could be a qualitative test for the functional state of DHP-sensitive Ca-channels.
Timofeev
A A
AA
Glebova
R N
RN
Kalinina
M V
MV
Grigorian
G L
GL
D000818:Animals; D002121:Calcium Channel Blockers / Q000032:analysis*; D015220:Calcium Channels / Q000378:metabolism*; D002540:Cerebral Cortex / Q000737:chemistry; D004095:Dihydropyridines / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D015335:Molecular Probes; D009543:Nifedipine / Q000031:analogs & derivatives* / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D013050:Spectrometry, Fluorescence; D013574:Synaptosomes / Q000737:chemistry*
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D004095:Dihydropyridines; D015335:Molecular Probes; D009543:Nifedipine; C045912:ryodipine
Ispol'zovanie spektral'nykh kharakteristik riodipina dlia izucheniia digidropiridin-retseptornogo kompleksa neĭronal'nykh membran.