5JBN
Crystal Structure of Apo Phosphopantetheine Adenylyltransferase (PPAT/CoaD) from E. coli
Summary for 5JBN
| Entry DOI | 10.2210/pdb5jbn/pdb |
| Descriptor | Phosphopantetheine adenylyltransferase, SULFATE ION, DIMETHYL SULFOXIDE, ... (4 entities in total) |
| Functional Keywords | coad, apo, ppat, transferase |
| Biological source | Escherichia coli (strain K12) |
| Cellular location | Cytoplasm: P0A6I6 |
| Total number of polymer chains | 2 |
| Total formula weight | 39146.76 |
| Authors | Mamo, M.,Proudfoot, A.,Bussiere, D. (deposition date: 2016-04-13, release date: 2016-05-25, Last modification date: 2024-03-06) |
| Primary citation | Proudfoot, A.,Frank, A.O.,Ruggiu, F.,Mamo, M.,Lingel, A. Facilitating unambiguous NMR assignments and enabling higher probe density through selective labeling of all methyl containing amino acids. J.Biomol.Nmr, 65:15-27, 2016 Cited by PubMed Abstract: The deuteration of proteins and selective labeling of side chain methyl groups has greatly enhanced the molecular weight range of proteins and protein complexes which can be studied using solution NMR spectroscopy. Protocols for the selective labeling of all six methyl group containing amino acids individually are available, however to date, only a maximum of five amino acids have been labeled simultaneously. Here, we describe a new methodology for the simultaneous, selective labeling of all six methyl containing amino acids using the 115 kDa homohexameric enzyme CoaD from E. coli as a model system. The utility of the labeling protocol is demonstrated by efficiently and unambiguously assigning all methyl groups in the enzymatic active site using a single 4D (13)C-resolved HMQC-NOESY-HMQC experiment, in conjunction with a crystal structure. Furthermore, the six fold labeled protein was employed to characterize the interaction between the substrate analogue (R)-pantetheine and CoaD by chemical shift perturbations, demonstrating the benefit of the increased probe density. PubMed: 27130242DOI: 10.1007/s10858-016-0032-2 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.45 Å) |
Structure validation
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