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PDBsum entry 1v10
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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The structure of rigidoporus lignosus laccase containing a full complement of copper ions, Reveals an asymmetrical arrangement for the t3 copper pair.
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Authors
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S.Garavaglia,
M.T.Cambria,
M.Miglio,
S.Ragusa,
V.Iacobazzi,
F.Palmieri,
C.D'Ambrosio,
A.Scaloni,
M.Rizzi.
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Ref.
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J Mol Biol, 2004,
342,
1519-1531.
[DOI no: ]
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PubMed id
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Abstract
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Laccase is a multicopper blue oxidase that couples the four-electron reduction
of oxygen with the oxidation of a broad range of organic substrates, including
phenols and arylamines. The enzyme is the object of intense biotechnological
research, due to its employment in bioremediation of soils and water as well as
in other biotechnological applications. We report here the cDNA and protein
sequences, the post-translational modifications, the crystallization and X-ray
structure determination of a laccase from the white-rot fungus Rigidoporus
lignosus. The amino acid residues sequence deduced from cDNA clearly identified
a pre-sequence of 21 residues representing the signal for extra-cellular
localization. Mass spectrometry analysis performed on the salvage enzyme,
confirmed the deduced sequence and precisely mapped two glycosylation sites at
Asn337 and Asn435, determining the nature of the bound glycosidic moieties. The
crystal structure was determined at 1.7A resolution from perfectly hemihedrally
twinned crystals, by molecular replacement technique. While the overall
structure closely resembled those reported for other fungal laccases, the
analysis of the T2/T3 trinuclear cluster revealed an unprecedented coordination
sphere for the T3 copper pair. No bridging oxygen ligand was present between the
two T3 copper ions, which were no longer symmetrically coordinated. The observed
structure could represent an intermediate along the process of four-electron
reduction of oxygen to water taking place at the trinuclear copper cluster.
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Figure 5.
Figure 5. Representation of the T1 and T2/T3 copper centers in RlLa. (A) Schematic representation of the T1 (Cu1)
copper center, reporting inter-atomic distances between relevant atoms. (B) Stereo view around the T1 center; residues
and copper ion are represented as ball-and-stick and sphere, respectively. (C) Schematic representation of the T2/T3 tri-
nuclear copper center, reporting inter-atomic distances between relevant atoms. (D) Stereo view for the T2/T3 copper
ions (Cu2, Cu3b and Cu3a) and their coordinating residues; copper ions and water molecules are depicted as green and
red spheres, respectively. Amino acid residues are drawn as ball-and-stick. Panel (B) and (C) were generated using
MOLSCRIPT.
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Figure 6.
Figure 6. Stereo view around the
T2/T3 center showing the electron
density of the Fo-F c difference
Fourier omit map (contouring
level is 4 sigma). Water molecules
and copper atoms are depicted as
red and green sphere, respectively.
The figure was generated using
BOBSCRIPT.
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The above figures are
reprinted
by permission from Elsevier:
J Mol Biol
(2004,
342,
1519-1531)
copyright 2004.
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Secondary reference #1
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Title
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Production, Purification, And properties of an extracellular laccase from rigidoporus lignosus.
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Authors
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M.Cambria,
A.Cambria,
S.Ragusa,
E.Rizzarelli.
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Ref.
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Protein Expr Purif, 2000,
18,
141-147.
[DOI no: ]
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PubMed id
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