4PYT

Crystal structure of a MurB family EP-UDP-N-acetylglucosamine reductase

4PYT の概要

• エントリーDOI: 10.2210/pdb4pyt/pdb
• 分子名称: UDP-N-acetylenolpyruvoylglucosamine reductase, FLAVIN-ADENINE DINUCLEOTIDE, MAGNESIUM ION, ... (5 entities in total)
• 機能のキーワード: murb/flavoprotein, oxidoreductase, fad binding
• 由来する生物種: unidentified
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34690.05

構造登録者

Cao, H.,Franz, L.,Sen, S.,Bingman, C.A.,Auldridge, M.,Steinmetz, E.,Mead, D.,Phillips Jr., G.N. (登録日: 2014-03-27, 公開日: 2014-05-21, 最終更新日: 2024-05-22)

主引用文献

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

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: 25906065
DOI: 10.1371/journal.pone.0124272

実験手法

X-RAY DIFFRACTION (1.853 Å)