Mutant insulin degrading enzyme and methods of use
AssigneeUniversity of Kentucky Research Foundation
InventorsDavid W. Rodgers, Louis B. Hersh, Nicholas Noinaj, Eun Suk Song
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Abstract
In one aspect, the present invention provides an isolated mutant insulin degrading enzyme (IDE) having an amino acid sequence that is at least 90% identical to SEQ ID NO:1 over its entire length and comprises at least one amino acid substitution at any of amino acid residues 332, 339, 341, 359, 360, 361, 374, 429, 609, 898, 899 or 901 of the sequence. The mutant IDE has a differential activity relative to that of wild-type IDE. Also provided is a polynucleotide encoding the polypeptide of the invention.
Claims19
An isolated mutant insulin degrading enzyme (IDE) comprising at least one amino acid substitution at any of amino acid residues 332, 339, 341, 359, 360, 361, 374, 429, 609, 898, 899 or 901 of SEQ ID NO:1 or SEQ ID NO:2;
- wherein the isolated mutant IDE has peptidase activity.
An isolated mutant insulin degrading enzyme (IDE) comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:1 over its entire length and comprises at least one amino acid substitution at any of amino acid residues 332, 339, 341, 359, 360, 361, 374, 429, 609, 898, 899 or 901 of the sequence;
- and wherein the isolated mutant IDE has peptidase activity.
A method of screening agents that modulate the activity of a mutant IDE;
- said method comprising the steps of (a) contacting a test agent with the mutant IDE in the presence of an IDE substrate, wherein the mutant IDE comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:1 over its entire length and comprises at least one amino acid substitution at any of amino acid residues 332, 339, 341, 359, 360, 361, 374, 429, 609, 898, 899 or 901 of the sequence;
- (b) assessing the effect of a test agent on activity of the mutant IDE, wherein activity is measured as effect of the test agent on the affinity of the mutant IDE for the IDE substrate or the test agent, rate of the IDE substrate cleavage, or stability of the mutant IDE, relative to a control lacking the test agent;
- and (c) selecting any test agent that modulates the affinity of the mutant IDE for the IDE substrate or the test agent, the rate of the IDE substrate cleavage, or the stability of the mutant IDE.
Dependencies are inferred from the claim text.