Complexome Profiling Identifies TMEM126B as a Component of the Mitochondrial Complex I Assembly Complex

Macromolecular complexes are essential players in numerous biological processes. They are often large, dynamic, and rather labile; approaches to study them are scarce. Covering masses up to ∼30 MDa, we separated the native complexome of rat heart mitochondria by blue-native and large-pore blue-nativ...

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Published in:Cell metabolism 2012-10, Vol.16 (4), p.538-549
Main Authors: Heide, Heinrich, Bleier, Lea, Steger, Mirco, Ackermann, Jörg, Dröse, Stefan, Schwamb, Bettina, Zörnig, Martin, Reichert, Andreas S., Koch, Ina, Wittig, Ilka, Brandt, Ulrich
Format: Article
Language:English
Subjects:
Animals
Databases, Protein
Electrophoresis, Gel, Two-Dimensional
HEK293 Cells
Humans
Male
Mass Spectrometry
Membrane Proteins - antagonists & inhibitors
Membrane Proteins - genetics
Membrane Proteins - metabolism
Mitochondria, Heart - metabolism
Multiprotein Complexes - chemistry
Multiprotein Complexes - metabolism
Proteomics
Rats
Rats, Wistar
RNA Interference
RNA, Small Interfering - metabolism
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Summary:Macromolecular complexes are essential players in numerous biological processes. They are often large, dynamic, and rather labile; approaches to study them are scarce. Covering masses up to ∼30 MDa, we separated the native complexome of rat heart mitochondria by blue-native and large-pore blue-native gel electrophoresis to analyze its constituents by mass spectrometry. Similarities in migration patterns allowed hierarchical clustering into interaction profiles representing a comprehensive analysis of soluble and membrane-bound complexes of an entire organelle. The power of this bottom-up approach was validated with well-characterized mitochondrial multiprotein complexes. TMEM126B was found to comigrate with known assembly factors of mitochondrial complex I, namely CIA30, Ecsit, and Acad9. We propose terming this complex mitochondrial complex I assembly (MCIA) complex. Furthermore, we demonstrate that TMEM126B is required for assembly of complex I. In summary, complexome profiling is a powerful and unbiased technique allowing the identification of previously overlooked components of large multiprotein complexes. [Display omitted] ► Multiprotein complexes up to a mass of 30 MDa are analyzed by proteomic profiling ► The power of complexome profiling is shown for mitochondrial multiprotein complexes ► TMEM126B is essential for the assembly of mitochondrial complex I ► TMEM126B is a subunit of the mitochondrial complex I assembly (MCIA) complex
ISSN:1550-4131
1932-7420
DOI:10.1016/j.cmet.2012.08.009