Nucleic acid-like proteins
Methods are provided for labeling and for detecting DNA-interacting proteins with an assayable label, comprising combining the DNA-interacting protein with a mfpA pentapeptide repeat family protein that has been purified and that binds to the protein, wherein the mfpA pentapeptide repeat family protein is bound to an assayable label.
1. A method of detecting a DNA-interacting protein, the method comprising
(a) contacting a protein with a purified Mycobacterium tuberculosis MfpA pentapeptide repeat protein (MtMfpA) which has had an assayable label bound thereto, wherein the label is a visible label, a fluorescent protein, a fluorescent organic compound less than 2000 molecular weight, a radioactive molecule, or an oligohistidine sequence, under conditions permitting the MtMfpA pentapeptide repeat protein to bind to a DNA-interacting protein, and
(b) detecting the visible label, fluorescent protein, fluorescent organic compound less than 2000 molecular weight, radioactive molecule, or oligohistidine sequence, respectively, of the labeled MtMfpA that is bound to the DNA-interacting protein, so as to thereby detect the DNA-interacting protein.
2. The method of claim 1 , wherein the assayable label is a fluorescent protein or a fluorescent organic compound of less than 2000 molecular weight.
3. The method of claim 1 , wherein the DNA-interacting protein is a DNA metabolizing or DNA catabolizing enzyme.
4. The method of claim 3 , wherein the DNA-metabolizing or DNA catabolizing enzyme is a DNA gyrase, a DNA polymerase, an RNA polymerase, a reverse transcriptase, a DNA ligase, an RNA ligase, a polynucleotide kinase, an alkaline phosphatase, a pyrophosphatase, a DNA glycosylase, a topoisomerase, a nicking enzyme, a restriction endonuclease, a ribonuclease, a recombinase, a deoxyribonuclease, or an exonuclease.
5. The method of claim 3 , wherein the DNA-metabolizing or DNA catabolizing enzyme is a DNA gyrase.
6. The method of claim 1 , wherein the DNA-interacting protein is a DNA-binding protein.
7. The method of claim 6 , wherein the DNA-binding protein is a single stranded DNA binding protein, a transcription factor, a repressor, an activator, an enhancer, a helix-turn-helix protein, a zinc finger protein, a leucine zipper protein, a helix-loop-helix protein, a steroid receptor, or a homeodomain protein.
8. The method of claim 1 , wherein the purified Mycobacterium tuberculosis MfpA is encoded by an Rv3361c gene.
9. The method of claim 1 , wherein the purified Mycobacterium tuberculosis MfpA is 184 amino acids in length.