4PYT
Crystal structure of a MurB family EP-UDP-N-acetylglucosamine reductase
Summary for 4PYT
| Entry DOI | 10.2210/pdb4pyt/pdb |
| Descriptor | UDP-N-acetylenolpyruvoylglucosamine reductase, FLAVIN-ADENINE DINUCLEOTIDE, MAGNESIUM ION, ... (5 entities in total) |
| Functional Keywords | murb/flavoprotein, oxidoreductase, fad binding |
| Biological source | unidentified |
| Total number of polymer chains | 1 |
| Total formula weight | 34690.05 |
| Authors | Cao, H.,Franz, L.,Sen, S.,Bingman, C.A.,Auldridge, M.,Steinmetz, E.,Mead, D.,Phillips Jr., G.N. (deposition date: 2014-03-27, release date: 2014-05-21, Last modification date: 2024-05-22) |
| Primary citation | Auldridge, M.E.,Cao, H.,Sen, S.,Franz, L.P.,Bingman, C.A.,Yennamalli, R.M.,Phillips, G.N.,Mead, D.,Steinmetz, E.J. LucY: A Versatile New Fluorescent Reporter Protein. Plos One, 10:e0124272-e0124272, 2015 Cited by PubMed Abstract: We report on the discovery, isolation, and use of a novel yellow fluorescent protein. Lucigen Yellow (LucY) binds one FAD molecule within its core, thus shielding it from water and maintaining its structure so that fluorescence is 10-fold higher than freely soluble FAD. LucY displays excitation and emission spectra characteristic of FAD, with 3 excitation peaks at 276 nm, 377 nm, and 460 nm and a single emission peak at 530 nm. These excitation and emission maxima provide the large Stokes shift beneficial to fluorescence experimentation. LucY belongs to the MurB family of UDP-N-acetylenolpyruvylglucosamine reductases. The high resolution crystal structure shows that in contrast to other structurally resolved MurB enzymes, LucY does not contain a potentially quenching aromatic residue near the FAD isoalloxazine ring, which may explain its increased fluorescence over related proteins. Using E. coli as a system in which to develop LucY as a reporter, we show that it is amenable to circular permutation and use as a reporter of protein-protein interaction. Fragmentation between its distinct domains renders LucY non-fluorescent, but fluorescence can be partially restored by fusion of the fragments to interacting protein domains. Thus, LucY may find application in Protein-fragment Complementation Assays for evaluating protein-protein interactions. PubMed: 25906065DOI: 10.1371/journal.pone.0124272 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.853 Å) |
Structure validation
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