4pyt
Crystal structure of a MurB family EP-UDP-N-acetylglucosamine reductaseCrystal structure of a MurB family EP-UDP-N-acetylglucosamine reductase
Structural highlights
FunctionMURB_BACLD Cell wall formation.[HAMAP-Rule:MF_00037] Publication Abstract from PubMedWe 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. LucY: A Versatile New Fluorescent Reporter Protein.,Auldridge ME, Cao H, Sen S, Franz LP, Bingman CA, Yennamalli RM, Phillips GN Jr, Mead D, Steinmetz EJ PLoS One. 2015 Apr 23;10(4):e0124272. doi: 10.1371/journal.pone.0124272., eCollection 2015. PMID:25906065[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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