PDBsum entry 4pyt

Oxidoreductase

PDB id: 4pyt

Contents

Protein chain

302 a.a.

Ligands

FAD

Metals

_CL ×2

_MG ×3

Waters ×346

PDB id:

4pyt

Name:

Oxidoreductase

Title:

Crystal structure of a murb family ep-udp-n-acetylglucosamine reductase

Structure:

Udp-n-acetylenolpyruvoylglucosamine reductase. Chain: a. Synonym: udp-n-acetylmuramate dehydrogenase. Engineered: yes

Source:

Unidentified. Organism_taxid: 32644. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.85Å R-factor: 0.205 R-free: 0.227

Authors:

H.Cao,L.Franz,S.Sen,C.A.Bingman,M.Auldridge,E.Steinmetz,D.Mead, G.N.Phillips Jr.

Key ref:

M.E.Auldridge et al. (2015). LucY: A Versatile New Fluorescent Reporter Protein. Plos One, 10, e0124272. PubMed id: 25906065 DOI: 10.1371/journal.pone.0124272

Date:

27-Mar-14 Release date: 21-May-14

Protein chain

Q65JX9  (MURB_BACLD) -  UDP-N-acetylenolpyruvoylglucosamine reductase from Bacillus licheniformis (strain ATCC 14580 / DSM 13 / JCM 2505 / CCUG 7422 / NBRC 12200 / NCIMB 9375 / NCTC 10341 / NRRL NRS-1264 / Gibson 46)

Seq: 303 a.a.

Struc: 302 a.a.*

* PDB and UniProt seqs differ at 21 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.1.3.1.98 - UDP-N-acetylmuramate dehydrogenase.
• Reaction: UDP-N-acetyl-alpha-D-muramate + NADP⁺ = UDP-N-acetyl-3- O-(1-carboxyvinyl)-alpha-D-glucosamine + NADPH + H⁺
• Cofactor: FAD

FAD (Bound ligand (Het Group name = FAD) corresponds exactly)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1371/journal.pone.0124272

Plos One 10:e0124272 (2015)

PubMed id: 25906065

LucY: A Versatile New Fluorescent Reporter Protein.

M.E.Auldridge, H.Cao, S.Sen, L.P.Franz, C.A.Bingman, R.M.Yennamalli, G.N.Phillips, D.Mead, E.J.Steinmetz.

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.