Methods for Assessing CDK5 Activation and Function
AssigneeSilverbrook Research Pty Ltd
InventorsJames A. Bibb, Ammar H. Hawasli
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Abstract
As described herein, signaling events occurring in neurons or at neuronal synapses have been identified that involve Cdk5 and various other molecules which bind to, are activated by, and/or activate Cdk5. Of particular relevance are interactions that stimulate calpain cleavage of p35 into p25, which binds Cdk5 in pathologic states. Assays to identify modulators of these interactions are provided.
Claims26
A method of identifying an inhibitor of Cdk5/p25 complex formation comprising:
- (a) providing a system that permits formation of Cdk5/p25 complex formation;
- (b) introducing into said system a candidate substance;
- and
- (c) assessing Cdk5/p25 complex formation using a p25 selective antibody,
- wherein a candidate substance that reduces Cdk5/p25 complex formation, as compared to Cdk5/p25 complex formation observed in the absence of said candidate substance, identifies said candidate substance as an inhibitor of Cdk5/p25 formation.
(canceled)
A method of identifying an inhibitor of calpain-dependent p25 formation comprising:
- (a) providing a system that permits cleavage of p35 into p25 by calpain;
- (b) introducing into said system a candidate substance;
- and
- (c) assessing p25 formation using a p25 selective antibody,
- wherein a candidate substance that reduces p25 formation, as compared to p25 formation observed in the absence of said candidate substance, identifies said candidate substance as an inhibitor of calpain-dependent p25 formation.
(canceled)
A method of identifying an inhibitor of calpain activation comprising:
- (a) providing a system comprising calpain and Cdk5;
- (b) introducing into said system a candidate substance;
- and
- (c) assessing Cdk5/caplain complex formation or calpain proteolytic activity,
- wherein a candidate substance that reduces Cdk5/calpain complex formation or calpain proteolytic activity, as compared to Cdk5/calpain complex formation or calpain proteolytic activity observed in the absence of said candidate substance, identifies said candidate substance as an inhibitor of calpain activation.
(canceled)
A method of identifying an inhibitor of Cdk5-NR2B complex formation comprising:
- (a) providing a system comprising NR2B and Cdk5;
- (b) introducing into said system a candidate substance;
- and
- (c) assessing Cdk5/NR2B complex formation or Cdk5 phosphorylation activity,
- wherein a candidate substance that reduces Cdk5/NR2B complex formation or reduces Cdk5 phosphorylation activity, as compared to Cdk5/NR2B complex formation or reduces Cdk5 phosphorylation activity observed in the absence of said candidate substance, identifies said candidate substance as an inhibitor of Cdk5-NR2B formation or reduces Cdk5 phosphorylation activity.
(canceled)
Dependencies are inferred from the claim text.