Protocol to characterize mitochondrial supercomplexes from mouse tissues by combining BN-PAGE and MS-based proteomics
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Protocol to characterize mitochondrial supercomplexes from mouse tissues by combining BN-PAGE and MS-based proteomics. / Moreno-Justicia, Roger; Gonzalez-Franquesa, Alba; Stocks, Ben; Deshmukh, Atul S.
In: STAR Protocols, Vol. 3, No. 1, 101135, 2022.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Protocol to characterize mitochondrial supercomplexes from mouse tissues by combining BN-PAGE and MS-based proteomics
AU - Moreno-Justicia, Roger
AU - Gonzalez-Franquesa, Alba
AU - Stocks, Ben
AU - Deshmukh, Atul S.
N1 - Publisher Copyright: © 2022 The Author(s)
PY - 2022
Y1 - 2022
N2 - The assembly of mitochondrial respiratory complexes into supercomplexes has significant implications for mitochondrial function. This protocol details mitochondrial isolation from mouse tissues and the use of blue native gel electrophoresis (BN-PAGE) to separate pre-identified mitochondrial supercomplexes into different gel bands. We then describe the excision of the individual bands, followed by in-gel protein digestion and peptide desalting for mass spectrometry (MS)-based proteomics. This protocol provides a time-efficient measurement of the abundance and distribution of proteins within known supercomplexes. For complete details on the use and execution of this profile, please refer to Gonzalez-Franquesa et al. (2021).
AB - The assembly of mitochondrial respiratory complexes into supercomplexes has significant implications for mitochondrial function. This protocol details mitochondrial isolation from mouse tissues and the use of blue native gel electrophoresis (BN-PAGE) to separate pre-identified mitochondrial supercomplexes into different gel bands. We then describe the excision of the individual bands, followed by in-gel protein digestion and peptide desalting for mass spectrometry (MS)-based proteomics. This protocol provides a time-efficient measurement of the abundance and distribution of proteins within known supercomplexes. For complete details on the use and execution of this profile, please refer to Gonzalez-Franquesa et al. (2021).
KW - Mass Spectrometry
KW - Metabolism
KW - Protein Biochemistry
KW - Proteomics
U2 - 10.1016/j.xpro.2022.101135
DO - 10.1016/j.xpro.2022.101135
M3 - Journal article
C2 - 35128478
AN - SCOPUS:85123698461
VL - 3
JO - STAR Protocols
JF - STAR Protocols
SN - 2666-1667
IS - 1
M1 - 101135
ER -
ID: 292000510