Details
| Original language | English |
|---|---|
| Pages (from-to) | 1079-1092 |
| Number of pages | 14 |
| Journal | Plant Journal |
| Volume | 89 |
| Issue number | 6 |
| Early online date | 10 Dec 2016 |
| Publication status | Published - 4 Mar 2017 |
Abstract
Mitochondria are central to cellular metabolism and energy conversion. In plants they also enable photosynthesis through additional components and functional flexibility. A majority of those processes relies on the assembly of individual proteins to larger protein complexes, some of which operate as large molecular machines. There has been a strong interest in the makeup and function of mitochondrial protein complexes and protein–protein interactions in plants, but the experimental approaches used typically suffer from selectivity or bias. Here, we present a complexome profiling analysis for leaf mitochondria of the model plant Arabidopsis thaliana for the systematic characterization of protein assemblies. Purified organelle extracts were separated by 1D Blue native (BN) PAGE, a resulting gel lane was dissected into 70 slices (complexome fractions) and proteins in each slice were identified by label free quantitative shot-gun proteomics. Overall, 1359 unique proteins were identified, which were, on average, present in 17 complexome fractions each. Quantitative profiles of proteins along the BN gel lane were aligned by similarity, allowing us to visualize protein assemblies. The data allow re-annotating the subunit compositions of OXPHOS complexes, identifying assembly intermediates of OXPHOS complexes and assemblies of alternative respiratory oxidoreductases. Several protein complexes were discovered that have not yet been reported in plants, such as a 530 kDa Tat complex, 460 and 1000 kDa SAM complexes, a calcium ion uniporter complex (150 kDa) and several PPR protein complexes. We have set up a tailored online resource (https://complexomemap.de/at_mito_leaves) to deposit the data and to allow straightforward access and custom data analyses.
Keywords
- alternative electron transport, Arabidopsis thaliana, complex I, mitochondrial calcium uniporter, mitochondrial metabolism, PPR proteins, proteomics, respiratory chain
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
- Agricultural and Biological Sciences(all)
- Plant Science
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Plant Journal, Vol. 89, No. 6, 04.03.2017, p. 1079-1092.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - The mitochondrial complexome of Arabidopsis thaliana
AU - Senkler, Jennifer
AU - Senkler, Michael
AU - Eubel, Holger
AU - Hildebrandt, Tatjana
AU - Lengwenus, Christian
AU - Schertl, Peter
AU - Schwarzländer, Markus
AU - Wagner, Stephan
AU - Wittig, Ilka
AU - Braun, Hans Peter
N1 - Funding information: This work was supported by the Deutsche Forschungsgemeinschaft (grant EU54/4-1 to HE, grant SCHW1719/1-1 to MS, grant SFB815/Z1 to IW and grant BR1829/10-2 to HPB).
PY - 2017/3/4
Y1 - 2017/3/4
N2 - Mitochondria are central to cellular metabolism and energy conversion. In plants they also enable photosynthesis through additional components and functional flexibility. A majority of those processes relies on the assembly of individual proteins to larger protein complexes, some of which operate as large molecular machines. There has been a strong interest in the makeup and function of mitochondrial protein complexes and protein–protein interactions in plants, but the experimental approaches used typically suffer from selectivity or bias. Here, we present a complexome profiling analysis for leaf mitochondria of the model plant Arabidopsis thaliana for the systematic characterization of protein assemblies. Purified organelle extracts were separated by 1D Blue native (BN) PAGE, a resulting gel lane was dissected into 70 slices (complexome fractions) and proteins in each slice were identified by label free quantitative shot-gun proteomics. Overall, 1359 unique proteins were identified, which were, on average, present in 17 complexome fractions each. Quantitative profiles of proteins along the BN gel lane were aligned by similarity, allowing us to visualize protein assemblies. The data allow re-annotating the subunit compositions of OXPHOS complexes, identifying assembly intermediates of OXPHOS complexes and assemblies of alternative respiratory oxidoreductases. Several protein complexes were discovered that have not yet been reported in plants, such as a 530 kDa Tat complex, 460 and 1000 kDa SAM complexes, a calcium ion uniporter complex (150 kDa) and several PPR protein complexes. We have set up a tailored online resource (https://complexomemap.de/at_mito_leaves) to deposit the data and to allow straightforward access and custom data analyses.
AB - Mitochondria are central to cellular metabolism and energy conversion. In plants they also enable photosynthesis through additional components and functional flexibility. A majority of those processes relies on the assembly of individual proteins to larger protein complexes, some of which operate as large molecular machines. There has been a strong interest in the makeup and function of mitochondrial protein complexes and protein–protein interactions in plants, but the experimental approaches used typically suffer from selectivity or bias. Here, we present a complexome profiling analysis for leaf mitochondria of the model plant Arabidopsis thaliana for the systematic characterization of protein assemblies. Purified organelle extracts were separated by 1D Blue native (BN) PAGE, a resulting gel lane was dissected into 70 slices (complexome fractions) and proteins in each slice were identified by label free quantitative shot-gun proteomics. Overall, 1359 unique proteins were identified, which were, on average, present in 17 complexome fractions each. Quantitative profiles of proteins along the BN gel lane were aligned by similarity, allowing us to visualize protein assemblies. The data allow re-annotating the subunit compositions of OXPHOS complexes, identifying assembly intermediates of OXPHOS complexes and assemblies of alternative respiratory oxidoreductases. Several protein complexes were discovered that have not yet been reported in plants, such as a 530 kDa Tat complex, 460 and 1000 kDa SAM complexes, a calcium ion uniporter complex (150 kDa) and several PPR protein complexes. We have set up a tailored online resource (https://complexomemap.de/at_mito_leaves) to deposit the data and to allow straightforward access and custom data analyses.
KW - alternative electron transport
KW - Arabidopsis thaliana
KW - complex I
KW - mitochondrial calcium uniporter
KW - mitochondrial metabolism
KW - PPR proteins
KW - proteomics
KW - respiratory chain
UR - http://www.scopus.com/inward/record.url?scp=85011429878&partnerID=8YFLogxK
U2 - 10.1111/tpj.13448
DO - 10.1111/tpj.13448
M3 - Article
C2 - 27943495
AN - SCOPUS:85011429878
VL - 89
SP - 1079
EP - 1092
JO - Plant Journal
JF - Plant Journal
SN - 0960-7412
IS - 6
ER -