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PDBsum entry 1j2v
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Structural genomics, unknown function
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PDB id
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1j2v
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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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Structural implications for heavy metal-Induced reversible assembly and aggregation of a protein: the case of pyrococcus horikoshii cuta.
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Authors
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Y.Tanaka,
K.Tsumoto,
T.Nakanishi,
Y.Yasutake,
N.Sakai,
M.Yao,
I.Tanaka,
I.Kumagai.
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Ref.
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FEBS Lett, 2004,
556,
167-174.
[DOI no: ]
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PubMed id
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Abstract
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CutA is a small protein that appears to be involved in the mechanism of divalent
metal cation tolerance in microorganisms. Here we report the crystal structure
of Pyrococcus horikoshii CutA (PhoCutA), with and without Cu(2+), and its
metal-binding properties. Crystallographic analyses revealed that PhoCutA forms
a stable trimeric structure with intertwined antiparallel beta-strands. The
crystal structure of the Cu(2+)-PhoCutA complex shows that the Cu(2+) is located
at a trimer-trimer interface and is recognized by the side chains of one Asp(48)
from each trimer. In an in vitro experiment, PhoCutA bound to several heavy
metals, most of which led to reversible aggregation of the protein; i.e. the
aggregates could be completely solubilized by addition of ethylenediamine
tetraacetic acid (EDTA) or dialysis against metal free buffer. Substitution of
Asp(48) with Ala led to a decrease in the amount of aggregates, suggesting the
significant contribution of Asp(48) to the reversible aggregation. To the best
of our knowledge, this is the first report which provides the structural
evidence for heavy metal-induced multimerization of a protein.
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Figure 3.
Fig. 3. Structure of PhoCutA complexed with copper. A:
Superposition of the Cα trace of PhoCutA without Cu^2+ (blue)
and complexed with Cu^2+ (red). Cu^2+ (green ball) and Asp^48
(ball-and-stick) are also shown. B: Ribbon diagram of dimer of
trimeric structures. Copper ion (Cu^2+) is shown as a green ball
in the trimer–trimer interface. C: Deduced structure of
multimer induced by Cu^2+ binding. Cu^2+ is green, proteins in
the first layer are blue, and proteins in the second layer are
red. D: Cu^2+-binding site of PhoCutA. Each trimer is colored
blue or red. Cu^2+-binding residues (ball-and-stick
representations) and water molecules (red balls) are shown.
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Figure 4.
Fig. 4. SDS–PAGE of PhoCutA untreated by metals.
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The above figures are
reprinted
by permission from the Federation of European Biochemical Societies:
FEBS Lett
(2004,
556,
167-174)
copyright 2004.
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