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Rationally designed helix-turn-helix proteins and their conformational changes upon DNA binding.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Percipalle, Piergiorgio Simoncsits, András Zakhariev, Sotir Guarnaccia, Corrado Sánchez, Ramiro Pongor, Sándor |
| Copyright Year | 1995 |
| Abstract | Circular dichroism and electrophoretic mobility shift studies were performed to confirm that dimerized N-terminal domains of bacterial repressors containing helix-turn-helix motifs are capable of high-affinity and specific DNA recognition as opposed to the monomeric N-terminal domains. Specific, high-affinity DNA binding proteins were designed and produced in which two copies of the N-terminal 1-62 domain of the bacteriophage 434 repressor are connected either in a dyad-symmetric fashion, with a synthetic linker attached to the C-termini, or as direct sequence repeats. Both molecules bound to their presumptive cognate nearly as tightly as does the natural (full-length and non-covalently dimerized) 434 repressor, showing that covalent dimerization can be used to greatly enhance the binding activity of individual protein segments. Circular dichroism spectroscopy showed a pronounced increase in the alpha-helix content when these new proteins interacted with their cognate DNA and a similar, although 30% lower, increase was also seen upon their interaction with non-cognate DNA. These results imply that a gradual conformational change may occur when helix-turn-helix motifs bind to DNA, and that a scanning mechanism is just as plausible for this motif class as that which is proposed for the more flexible basic-leucine zipper and basic-helix-loop-helix motifs. |
| File Format | PDF HTM / HTML |
| DOI | 10.1002/j.1460-2075.1995.tb07322.x |
| PubMed reference number | 7621832 |
| Journal | Medline |
| Volume Number | 14 |
| Issue Number | 13 |
| Alternate Webpage(s) | http://pongor.itk.ppke.hu/library/Group-Publications/PAPERS%20PDF%20ONLY/Piergiorgio%20Percipalle.pdf |
| Alternate Webpage(s) | https://doi.org/10.1002/j.1460-2075.1995.tb07322.x |
| Journal | The EMBO journal |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
