Mitogenic oxygenase regulators
The present invention relates to new genes encoding for the production of novel nox enzyme proteins involved in generation of reactive oxygen intermediates that affect cell division. The present invention also provides vectors containing these genes, cells transfected with these vectors, antibodies raised against these novel proteins, kits for detection, localization and measurement of these genes and proteins, and methods to determine the activity of drugs to affect the activity of the proteins of the present invention.
1. A method of determining the activity of an agent to decrease cell division, comprising contacting a protein comprising the amino acid sequence of SEQ ID NO:4, or a fragment of SEQ ID NO: 4 capable of superoxide production, or a substitution thereof of less than 1% of the amino acid sequence, with the agent; and measuring the ability of the agent to decrease superoxide production, inhibit diaphorase activity or bind the protein as compared to a control not contacted with the agent, thereby determining the ability of the agent to decrease cell division.
2. The method of claim 1 , wherein the protein comprises the amino acid sequence of SEQ ID NO:4 or a fragment of SEQ ID NO: 4 capable of superoxide production.
3. The method of claim 1 , wherein the method comprises measuring the ability of the agent to inhibit superoxide production.
4. The method of claim 1 , wherein the method comprises measuring the ability of the agent to bind the protein.
5. The method of claim 1 , wherein the protein comprises the C-terminal 265 amino acids of SEQ ID NO: 4.
6. The method of claim 1 , wherein the method comprises measuring the ability of the agent to inhibit diaphorase activity.
7. The method of claim 6 , wherein measuring the ability of the agent to inhibit the diaphorase activity comprises evaluating NAPDH-dependent reductase activity.
8. The method of claim 6 , wherein evaluating the diaphorase activity of the protein comprises use of cytochrome c or nitrobluetetrazolium.
9. A method for identifying an agent that modulates the activity of a Nox enzyme, comprising contacting an agent of interest with a protein comprising the C-terminal 265 amino acids of SEQ ID NO: 4 capable of superoxide production; and evaluating the diaphorase activity of the protein; wherein a decrease in the diaphorase activity of the protein as compared to a control indicates that the agent modulates the activity of a Nox enzyme.
10. The method of claim 9 , wherein evaluating the diaphorase activity of the protein comprises evaluating NAPDH-dependent reductase activity.
11. The method of claim 9 , wherein evaluating the diaphorase activity of the protein comprises use of cytochrorue c or nitrobluetetrazolium.