A proteomic screen reveals the mitochondrial outer membrane protein Mdm34p as an essential target of the F-box protein Mdm30p

Kazuhisa Ota, Keiji Kito, Satoshi Okada, Takashi Ito

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Ubiquitination plays various critical roles in eukaryotic cellular regulation and is medated by a cascade of enzymes including ubiquitin protein ligase (E3). The Skp1-Cullin-F-box protein complex comprises the largest E3 family, in each member of which a unique F-box protein binds its targets to define substrate specificity. Although genome sequencing uncovers a growing number of F-box proteins, most of them have remained as "orphans" because of the difficulties in identification of their substrates. To address this issue, we tested a quantitative proteomic approach by combining the stable isotope labeling by amino acids in cell culture (SILAC), parallel affinity purification (PAP) that we had developed for efficient enrichment of ubiquitinated proteins, and mass spectrometry (MS). We applied this SILAC-PAP-MS approach to compare ubiquitinated proteins between yeast cells with and without over-expressed Mdm30p, an F-box protein implicated in mitochondrial morphology. Consequently, we identified the mitochondrial outer membrane protein Mdm34p as a target of Mdm30p. Furthermore, we found that mitochondrial defects induced by deletion of MDM30 are not only recapitulated by a mutant Mdm34p defective in interaction with Mdm30p but alleviated by ubiquitination-mimicking forms of Mdm34p. These results indicate that Mdm34p is a physiologically important target of Mdm30p.

Original languageEnglish
Pages (from-to)1075-1085
Number of pages11
JournalGenes to Cells
Volume13
Issue number10
DOIs
Publication statusPublished - Dec 1 2008

Fingerprint

F-Box Proteins
Ubiquitinated Proteins
Proteomics
Membrane Proteins
Ubiquitination
Mass Spectrometry
SKP Cullin F-Box Protein Ligases
Isotope Labeling
Ubiquitin-Protein Ligases
Substrate Specificity
Cell Culture Techniques
Yeasts
Genome
Amino Acids
Enzymes

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

Cite this

A proteomic screen reveals the mitochondrial outer membrane protein Mdm34p as an essential target of the F-box protein Mdm30p. / Ota, Kazuhisa; Kito, Keiji; Okada, Satoshi; Ito, Takashi.

In: Genes to Cells, Vol. 13, No. 10, 01.12.2008, p. 1075-1085.

Research output: Contribution to journalArticle

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