Nematode MDH-like sequences
Disclosed are two nucleic acid molecules from M. incognita encoding malate dehydrogenase-like (MDH-like) polypeptides. The MDH-like polypeptide sequences are also provided, as are vectors, host cells, and recombinant methods for production of MDH-like nucleotides and polypeptides. The invention further relates to screening methods for identifying inhibitors and/or activators, as well as methods for antibody production.
1. A method of screening for a compound that alters the malate dehydrogenase activity of a polypeptide, the method comprising:
providing a polypeptide comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3 and has malate dehydrogenase activity;
contacting the polypeptide with a test compound; and
measuring the malate dehydrogenase activity of the polypeptide, wherein a change in the malate dehydrogenase activity of the polypeptide relative to the malate dehydrogenase activity of the polypeptide in the absence of the test compound is an indication that the test compound alters the malate dehydrogenase activity of the polypeptide.
2. A method for identifying a compound that alters the malate dehydrogenase activity of a polypeptide comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3 and has malate dehydrogenase activity the method comprising:
providing a first polypeptide comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3 and has malate dehydrogenase activity;
measuring the inhibition of the malate dehydrogenase activity of the first polypeptide by a test compound under a set of assay conditions,
providing a second polypeptide, wherein the second polypeptide is produced by a plant or mammal and has malate dehydrogenase activity; measuring the inhibition of the malate dehydrogenase activity of the second polypeptide by said test compound under the set of assay conditions, wherein a test compound that inhibits the malate dehydrogenase activity of the first polypeptide to a greater extent than the activity of the second polypeptide under the same set of assay conditions is a compound that alters the malate dehydrogenase activity of the polypeptide.
3. The method of claim 1 wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:3.
4. The method of claim 1 wherein the step of measuring malate dehydrogenase activity comprises measuring the conversion of malate to oxaloacetate.
5. The method of claim 1 wherein the step of measuring malate dehydrogenase activity comprises measuring the conversion of oxaloacetate to malate.
6. The method of claim 2 wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:3.
7. The method of claim 2 wherein the step of measuring malate dehydrogenase activity comprises measuring the conversion of malate to oxaloacetate.
8. The method of claim 2 wherein the step of measuring malate dehydrogenase activity comprises measuring the conversion of oxaloacetate to malate.
9. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO:3.
10. The method of claim 2 , wherein the first polypeptide comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO:3.