TY - JOUR
T1 - Structural implications for heavy metal-induced reversible assembly and aggregation of a protein
T2 - The case of Pyrococcus horikoshii CutA
AU - Tanaka, Yoshikazu
AU - Tsumoto, Kouhei
AU - Nakanishi, Takeshi
AU - Yasutake, Yoshiaki
AU - Sakai, Naoki
AU - Yao, Min
AU - Tanaka, Isao
AU - Kumagai, Izumi
N1 - Funding Information:
This work was supported in part by the National Project on Protein Structural and Functional Analyses from the Ministry of Education, Culture, Sports, Science and Technology of Japan. It was also partially supported by the Ministry of Education, Culture, Sports, Science and Technology by a Grant-in-Aid for the COE project, Giant Molecules and Complex Systems, 2002. We thank M. Kawamoto, Dr. H. Sakai, and Dr. K. Miura of the Japan Synchrotron Radiation Research Institute (JASRI) for their help with the X-ray diffraction experiments at SPring-8.
PY - 2004/1/2
Y1 - 2004/1/2
N2 - 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 β-strands. The crystal structure of the Cu2+- PhoCutA complex shows that the Cu2+ is located at a trimer-trimer interface and is recognized by the side chains of one Asp48 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 Asp48 with Ala led to a decrease in the amount of aggregates, suggesting the significant contribution of Asp48 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.
AB - 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 β-strands. The crystal structure of the Cu2+- PhoCutA complex shows that the Cu2+ is located at a trimer-trimer interface and is recognized by the side chains of one Asp48 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 Asp48 with Ala led to a decrease in the amount of aggregates, suggesting the significant contribution of Asp48 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.
KW - Crystal structure
KW - CutA
KW - Heavy metals
KW - Multimerization
KW - Pyrococcus horikoshii
KW - Reversible aggregation
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U2 - 10.1016/S0014-5793(03)01402-9
DO - 10.1016/S0014-5793(03)01402-9
M3 - Article
C2 - 14706845
AN - SCOPUS:0347985688
SN - 0014-5793
VL - 556
SP - 167
EP - 174
JO - FEBS Letters
JF - FEBS Letters
IS - 1-3
ER -