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PDBsum entry 5ebu
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Oxidoreductase
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PDB id
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5ebu
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PDB id:
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| Name: |
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Oxidoreductase
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Title:
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Aerococcus viridans l-lactate oxidase y215f mutant
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Structure:
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L-lactate oxidase. Chain: a, b, c, d, e, f, g, h. Engineered: yes. Mutation: yes
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Source:
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Aerococcus viridans. Organism_taxid: 1377. Expressed in: escherichia coli. Expression_system_taxid: 562.
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Resolution:
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2.60Å
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R-factor:
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0.184
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R-free:
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0.246
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Authors:
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D.Rainer,B.Nidetzky,D.K.Wilson
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Key ref:
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T.Stoisser
et al.
(2016).
Conformational flexibility related to enzyme activity: evidence for a dynamic active-site gatekeeper function of Tyr(215) in Aerococcus viridans lactate oxidase.
Sci Rep,
6,
27892.
PubMed id:
DOI:
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Date:
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19-Oct-15
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Release date:
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29-Jun-16
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PROCHECK
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Headers
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References
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Q44467
(LOX_AERVM) -
L-lactate oxidase from Aerococcus viridans (strain ATCC 11563 / DSM 20340 / CCUG 4311 / JCM 20461 / NBRC 12219 / NCTC 8251 / M1)
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Seq:
Struc:
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374 a.a.
367 a.a.*
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Key: |
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PfamA domain |
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Secondary structure |
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CATH domain |
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*
PDB and UniProt seqs differ
at 5 residue positions (black
crosses)
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DOI no:
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Sci Rep
6:27892
(2016)
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PubMed id:
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Conformational flexibility related to enzyme activity: evidence for a dynamic active-site gatekeeper function of Tyr(215) in Aerococcus viridans lactate oxidase.
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T.Stoisser,
M.Brunsteiner,
D.K.Wilson,
B.Nidetzky.
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ABSTRACT
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L-Lactate oxidase (LOX) belongs to a large family of flavoenzymes that catalyze
oxidation of α-hydroxy acids. How in these enzymes the protein structure
controls reactivity presents an important but elusive problem. LOX contains a
prominent tyrosine in the substrate binding pocket (Tyr(215) in Aerococcus
viridans LOX) that is partially responsible for securing a flexible loop which
sequesters the active site. To characterize the role of Tyr(215), effects of
substitutions of the tyrosine (Y215F, Y215H) were analyzed kinetically,
crystallographically and by molecular dynamics simulations. Enzyme variants
showed slowed flavin reduction and oxidation by up to 33-fold. Pyruvate release
was also decelerated and in Y215F, it was the slowest step overall. A 2.6-Å
crystal structure of Y215F in complex with pyruvate shows the hydrogen bond
between the phenolic hydroxyl and the keto oxygen in pyruvate is replaced with a
potentially stronger hydrophobic interaction between the phenylalanine and the
methyl group of pyruvate. Residues 200 through 215 or 216 appear to be
disordered in two of the eight monomers in the asymmetric unit suggesting that
they function as a lid controlling substrate entry and product exit from the
active site. Substitutions of Tyr(215) can thus lead to a kinetic bottleneck in
product release.
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Links
