Metallothionein transfers zinc to mitochondrial aconitase through a direct interaction in mouse hearts

Wenke Feng, Jian Cai, William M. Pierce, Renty B. Franklin, Wolfgang Maret, Frederick W. Benz, Yujian Kang

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

Previous studies have shown that in a cell-free system, metallothionein (MT) releases zinc when the environment becomes oxidized and the released zinc is transferred to a zinc-binding protein if such a protein is present. However, it is unknown whether and how zinc transfers from MT to other proteins in vivo. The present study was undertaken to test the hypothesis that if zinc transfer from MT to other proteins occurs in vivo, the transfer would proceed through a direct interaction between MT and a specific group of proteins. The heart extract obtained from MT-null mice was incubated with 65Zn-MT or 65ZnCl2 and the proteins receiving 65Zn were separated by blue-native PAGE (BN-PAGE) or sodium dodecyl sulfate-PAGE (SDS-PAGE), and detected by autoradiography. A unique 65Zn-binding band was observed from the 65Zn-MT-incubated, but not the 65ZnCl2-incubated preparation. The analysis using matrix assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry revealed that mitochondrial aconitase (m-aconitase) was among the proteins accepting Zn directly from Zn-MT. The m-aconitase, not the cytosolic aconitase (c-aconitase), was co-immunoprecipitated with MT. This study demonstrates that MT transfers zinc to m-aconitase through a direct interaction.

Original languageEnglish (US)
Pages (from-to)853-858
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume332
Issue number3
DOIs
StatePublished - Jul 8 2005
Externally publishedYes

Fingerprint

Aconitate Hydratase
Metallothionein
Zinc
Proteins
Native Polyacrylamide Gel Electrophoresis
Cell-Free System
Autoradiography
zinc thionein
Polyacrylamide Gel Electrophoresis
Mass Spectrometry
Lasers
Sodium Dodecyl Sulfate
Ionization
Mass spectrometry
Desorption

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Metallothionein transfers zinc to mitochondrial aconitase through a direct interaction in mouse hearts. / Feng, Wenke; Cai, Jian; Pierce, William M.; Franklin, Renty B.; Maret, Wolfgang; Benz, Frederick W.; Kang, Yujian.

In: Biochemical and Biophysical Research Communications, Vol. 332, No. 3, 08.07.2005, p. 853-858.

Research output: Contribution to journalArticle

Feng, Wenke ; Cai, Jian ; Pierce, William M. ; Franklin, Renty B. ; Maret, Wolfgang ; Benz, Frederick W. ; Kang, Yujian. / Metallothionein transfers zinc to mitochondrial aconitase through a direct interaction in mouse hearts. In: Biochemical and Biophysical Research Communications. 2005 ; Vol. 332, No. 3. pp. 853-858.
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