4pyt: Difference between revisions
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4pyt FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4pyt OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4pyt RCSB], [http://www.ebi.ac.uk/pdbsum/4pyt PDBsum]</span></td></tr> | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4pyt FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4pyt OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4pyt RCSB], [http://www.ebi.ac.uk/pdbsum/4pyt PDBsum]</span></td></tr> | ||
</table> | </table> | ||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
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. | |||
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<ref>PMID:25906065</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
Revision as of 11:48, 12 August 2015
Crystal structure of a MurB family EP-UDP-N-acetylglucosamine reductaseCrystal structure of a MurB family EP-UDP-N-acetylglucosamine reductase
Structural highlights
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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