3PKP
Q83S Variant of S. Enterica RmlA with dATP
Summary for 3PKP
| Entry DOI | 10.2210/pdb3pkp/pdb |
| Related | 1IIM 1IIN 1MP3 1MP4 1MP5 3PKQ |
| Descriptor | Glucose-1-phosphate thymidylyltransferase, 2'-DEOXYADENOSINE 5'-TRIPHOSPHATE, MAGNESIUM ION, ... (4 entities in total) |
| Functional Keywords | nucleotidylyltransferase, directed evolution, structural genomics, psi-2, protein structure initiative, center for eukaryotic structural genomics, cesg, transferase |
| Biological source | Salmonella enterica subsp. enterica serovar Typhimurium |
| Total number of polymer chains | 8 |
| Total formula weight | 263879.94 |
| Authors | Chang, A.,Moretti, R.,Bingman, C.A.,Thorson, J.S.,Phillips Jr., G.N.,Center for Eukaryotic Structural Genomics (CESG) (deposition date: 2010-11-11, release date: 2011-01-12, Last modification date: 2024-02-21) |
| Primary citation | Moretti, R.,Chang, A.,Peltier-Pain, P.,Bingman, C.A.,Phillips, G.N.,Thorson, J.S. Expanding the Nucleotide and Sugar 1-Phosphate Promiscuity of Nucleotidyltransferase RmlA via Directed Evolution. J.Biol.Chem., 286:13235-13243, 2011 Cited by PubMed Abstract: Directed evolution is a valuable technique to improve enzyme activity in the absence of a priori structural knowledge, which can be typically enhanced via structure-guided strategies. In this study, a combination of both whole-gene error-prone polymerase chain reaction and site-saturation mutagenesis enabled the rapid identification of mutations that improved RmlA activity toward non-native substrates. These mutations have been shown to improve activities over 10-fold for several targeted substrates, including non-native pyrimidine- and purine-based NTPs as well as non-native D- and L-sugars (both α- and β-isomers). This study highlights the first broadly applicable high throughput sugar-1-phosphate nucleotidyltransferase screen and the first proof of concept for the directed evolution of this enzyme class toward the identification of uniquely permissive RmlA variants. PubMed: 21317292DOI: 10.1074/jbc.M110.206433 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.6 Å) |
Structure validation
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