LucY: A Versatile New Fluorescent Reporter Protein
dc.citation.firstpage | e0124272 | |
dc.citation.issueNumber | 4 | |
dc.citation.journalTitle | PLoS ONE | |
dc.citation.volumeNumber | 10 | |
dc.contributor.author | Auldridge, Michele E. | |
dc.contributor.author | Cao, Hongnan | |
dc.contributor.author | Sen, Saurabh | |
dc.contributor.author | Franz, Laura P. | |
dc.contributor.author | Bingman, Craig A. | |
dc.contributor.author | Yennamalli, Ragothaman M. | |
dc.contributor.author | Phillips, George N.Jr. | |
dc.contributor.author | Mead, David | |
dc.contributor.author | Steinmetz, Eric J. | |
dc.date.accessioned | 2015-11-09T18:52:18Z | |
dc.date.available | 2015-11-09T18:52:18Z | |
dc.date.issued | 2015 | |
dc.description.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. | |
dc.identifier.citation | Auldridge, Michele E., Cao, Hongnan, Sen, Saurabh, et al.. "LucY: A Versatile New Fluorescent Reporter Protein." <i>PLoS ONE,</i> 10, no. 4 (2015) Public Library of Science: e0124272. http://dx.doi.org/10.1371/journal.pone.0124272. | |
dc.identifier.doi | http://dx.doi.org/10.1371/journal.pone.0124272 | |
dc.identifier.uri | https://hdl.handle.net/1911/82039 | |
dc.language.iso | eng | |
dc.publisher | Public Library of Science | |
dc.rights | This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited | |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.title | LucY: A Versatile New Fluorescent Reporter Protein | |
dc.type | Journal article | |
dc.type.dcmi | Text | |
dc.type.publication | publisher version |
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