 |
PDBsum entry 4pyt
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Oxidoreductase
|
PDB id
|
|
|
|
4pyt
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Lucy: a versatile new fluorescent reporter protein.
|
|
|
Authors
|
|
M.E.Auldridge,
H.Cao,
S.Sen,
L.P.Franz,
C.A.Bingman,
R.M.Yennamalli,
G.N.Phillips,
D.Mead,
E.J.Steinmetz.
|
|
|
Ref.
|
|
Plos One, 2015,
10,
e0124272.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
|
|
|
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.
|
|
|
|
|
|
|
