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AssigneeUniversity of Iowa Research Foundation
InventorsKevin P. Campbell, Derrick R. Witcher, Peter McPherson, Steven D. Kahl, John D. Windass, Terence Lewis, Philip Bentley
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Disclosed is a novel photo-activatable derivative of ryanodine 9-hydroxy-21-(4-azidobenzoyloxy)-9-epiryanodine. This novel ryanodine derivative has been conjugated to keyhole limpet hemocyanin (KLH) and used for the production of antibodies with high affinity and specificity to ryanodine. A radioimmunoassay specific for ryanodine was developed using the anti-ryanodine antibodies, and a dissociation constant for ryanodine of 1 nM was determined. Also disclosed is the binding of a labeled, photoactivatable derivative of ryanodine (ABRy) with the ryanodine receptor of skeletal, cardiac and brain membranes. Digestion of the labeled ryanodine receptor revealed a labeled 76 kDa tryptic fragment which is critical for the formation of the high affinity ryanodine binding site.
An immunogenic derivative of ryanodine which, following immunization of a mammal, stimulates the production of antibodies which bind specifically to ryanodine with an IC.sub.50 of less than 10.sup.-8, the immunogenic derivative of ryanodine being characterized by the following structure:
A labeled affinity reagent which binds to the ryanodine receptor with an IC.sub.50 of less than about 10.sup.-6, the labeled affinity reagent being selected from the group consisting of labeled forms of 21-(2-[3,3,3-Trifluoro-2-diazopropionyloxy]-ethylmercapto)-ryanodine and 21-(4-Hydroxybutylmercapto)-ryanodine.
A labeled affinity reagent which binds to the ryanodine receptor with an IC.sub.50 of less than about 10.sup.-7, the labeled affinity reagent being selected from the group consisting of labeled forms of 10-O-(3-[4-Azidobenzamido]-propionyl)-ryanodine;
A method for identifying the ryanodine binding site on the ryanodine receptor, comprising:
Dependencies are inferred from the claim text.