Patent ID: 6168926
Filing Date: 2001-01-02
Classification: C07K,C12Q,G01N,Y10S

Abstract:
A method of screening test compounds for the ability to decrease or augment neuronal nitric oxide synthase (nNOS) activity, comprising the steps of: (a) contacting a cell with a test compound, wherein the cell comprises: i) a first fusion protein comprising (1) a DNA binding domain and (2) a mammalian protein inhibitor of nNOS (PIN-1) protein as shown in SEQ ID NO:2; ii) a second fusion protein comprising (1) a transcriptional activating domain and (2) amino acids 163-245 of a nNOS protein as shown in SEQ ID NO:3 or a domain of a nNOS protein as shown in SEQ ID NO:4 which binds to the PIN-1 protein, wherein the interaction of the PIN-1 protein with amino acids 163-245 of the nNOS protein as shown in SEQ ID NO:3 or with the domain of the nNOS protein as shown in SEQ ID NO:4 reconstitutes a sequence-specific transcriptional activating factor; and iii) a reporter gene comprising a DNA sequence to which the DNA binding domain of the first fusion protein specifically binds; and (b) measuring the expression of the reporter gene; wherein a test compound that increases the expression of the reporter gene would be a potential drug for decreasing nNOS activity, and a test compound that decreases the expression of the reporter gene would be a potential drug for augmenting nNOS activity.