| First Authors | Maria Luisa Romero Romero |
|---|---|
| Authors | Maria Luisa Romero Romero, Fan Yang, Yu-Ru Lin, Agnes Toth-Petroczy, Igor N. Berezovsky, Alexander Goncearenco, Wen Yang, Alon Wellner, Fanindra Kumar-Deshmukh, Michal Sharon, David Baker, Gabriele Varani, Dan S Tawfik |
| Corresponding Authors | |
| Last Authors | Dan S Tawfik |
| Journal Name | Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U.S.A.) |
| Volume | 115 |
| Issue | 51 |
| Page Range | 11943-11950 |
| PubMed ID | 30504143 |
| WebOfScience Link | WOS:000453529800010 |
| Open Access | false |
| Print Publication Date | 2018-12-18 |
| Online Publication Date | |
| Abstract | Abundant and essential motifs, such as phosphate-binding loops (P-loops), are presumed to be the seeds of modern enzymes. The Walker-A P-loop is absolutely essential in modern NTPase enzymes, in mediating binding, and transfer of the terminal phosphate groups of NTPs. However, NTPase function depends on many additional active-site residues placed throughout the protein's scaffold. Can motifs such as P-loops confer function in a simpler context? We applied a phylogenetic analysis that yielded a sequence logo of the putative ancestral Walker-A P-loop element: a β-strand connected to an α-helix via the P-loop. Computational design incorporated this element into de novo designed β-α repeat proteins with relatively few sequence modifications. We obtained soluble, stable proteins that unlike modern P-loop NTPases bound ATP in a magnesium-independent manner. Foremost, these simple P-loop proteins avidly bound polynucleotides, RNA, and single-strand DNA, and mutations in the P-loop's key residues abolished binding. Binding appears to be facilitated by the structural plasticity of these proteins, including quaternary structure polymorphism that promotes a combined action of multiple P-loops. Accordingly, oligomerization enabled a 55-aa protein carrying a single P-loop to confer avid polynucleotide binding. Overall, our results show that the P-loop Walker-A motif can be implemented in small and simple β-α repeat proteins, primarily as a polynucleotide binding motif. |
| Cover Image | |
| Affiliated With | Tóth-Petróczy |
| Selected By | |
| Acknowledged Services | |
| Publication Status | Published |
| Edoc Link | |
| Sfx Link | |
| DOI | 10.1073/pnas.1812400115 |
| Display Publisher Download Only | false |
| Visible On MPI-CBG Website | true |
| PDF Downloadable | true |
| Created By | verhegge |
| Added Date | 2018-11-26 |
| Last Edited By | verhegge |
| Last Edited Date | 2019-01-07 15:21:58.985 |
| Library ID | 7273 |
| Document ID | |
| Entry Complete | true |
| eDoc Compliant | true |
| Include in Edoc Report | true |
| Author Affiliations Complete | false |