Structural implications for heavy metal-induced reversible assembly and aggregation of a protein: The case of Pyrococcus horikoshii CutA

Yoshikazu Tanaka, Kouhei Tsumoto, Takeshi Nakanishi, Yoshiaki Yasutake, Naoki Sakai, Min Yao, Isao Tanaka, Izumi Kumagai

Research output: Contribution to journalArticlepeer-review

40 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)167-174
Number of pages8
JournalFEBS Letters
Volume556
Issue number1-3
DOIs
Publication statusPublished - 2004 Jan 2

Keywords

  • Crystal structure
  • CutA
  • Heavy metals
  • Multimerization
  • Pyrococcus horikoshii
  • Reversible aggregation

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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