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A single mitochondrion is a bean-shaped organelle enclosed by a double-membrane system. The outer membrane of mitochondria is smooth and contains many porins - the integral membrane transporters. Porins enable free diffusion of ions and small uncharged molecules through the outer mitochondrial membrane but limit the transport of molecules larger than 5000 Daltons. Further, the outer mitochondrial membrane forms a unique structure called membrane contact sites with other subcellular organelles,...
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Mitochondria are double-membrane organelles of the eukaryotes involved in cellular metabolism, signaling, ATP synthesis, and programmed cell death.  Each of these processes requires specific proteins and enzymes that must be correctly sorted to the right mitochondrial subcompartment for the proper functioning of the organelle.
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Mitochondria are eukaryotic cellular organelles that are known to produce energy through a process called oxidative phosphorylation. Besides their primary function, mitochondria are involved in various cellular processes, including cell growth, differentiation, signaling, metabolism, and senescence. Age-related changes cause a decline in mitochondrial quality and integrity due to increased mitochondrial mutations and oxidative damage. Thus, aging can severely impact mitochondrial functions,...
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Mitochondrial precursors are partially unfolded or loosely folded polypeptide chains. Newly synthesized precursors are inhibited from spontaneously folding into their native conformation by the cytosolic chaperones, heat shock proteins 70 (Hsp70), and mitochondrial import stimulation factors (MSFs). Precursors bound to MSFs are guided to the TOM70-TOM37 receptors, while precursors bound to Hsp70  chaperones are targetted to TOM20-TOM22 receptor complexes.
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The present-day mitochondrial and chloroplast genomes have retained some of the characteristics of their ancestral prokaryotes and also have acquired new attributes during their evolution within eukaryotic cells. Like prokaryotic genomes, mitochondrial and chloroplast genomes neither bind with histone-like proteins nor show complex packaging into chromosome-like structures, as observed in eukaryotes. Unlike mitotic cell divisions observed in eukaryotic cells, mitochondria and chloroplasts...
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The mitochondrial electron transport chain (ETC) is the main energy generation system in the eukaryotic cells. However, mitochondria also produce cytotoxic reactive oxygen species (ROS) due to the large electron flow during oxidative phosphorylation. While Complex I is one of the primary sources of superoxide radicals, ROS production by Complex II is uncommon and may only be observed in cancer cells with mutated complexes.
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High-Resolution Complexome Profiling by Cryoslicing BN-MS Analysis
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Mitochondrial Complexome Profiling.

Heiko Giese1, Jana Meisterknecht2, Juliana Heidler2

  • 1Molecular Bioinformatics, Institute of Computer Science, Goethe-University, Frankfurt am Main, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|November 24, 2020
PubMed
Summary
This summary is machine-generated.

Complexome profiling, using blue native gel electrophoresis and mass spectrometry, identifies protein interactions and assembly dynamics. This powerful technique aids in understanding cellular and disease mechanisms.

Keywords:
AssemblyBlue native electrophoresisComplexome profilingMass spectrometryMembrane protein complexesMitochondria

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Area of Science:

  • Biochemistry and Molecular Biology
  • Proteomics
  • Cell Biology

Background:

  • Understanding cellular protein interactions is crucial for deciphering biological processes.
  • Existing methods may lack the sensitivity to detect low-abundance protein assemblies.
  • Dynamic changes in protein complex formation are key to cellular function and dysfunction.

Purpose of the Study:

  • To describe the comprehensive workflow of complexome profiling.
  • To highlight the utility of complexome profiling in identifying protein interactomes.
  • To showcase its application in studying dynamic protein assembly processes.

Main Methods:

  • Combines blue native gel electrophoresis (BNE) with quantitative mass spectrometry.
  • Enables the separation and identification of intact protein complexes.
  • Integrates data analysis using specialized bioinformatics tools.

Main Results:

  • Successfully defines the entire protein interactome of cells, organelles, or membrane preparations.
  • Identifies protein assemblies, including those with low abundance.
  • Detects dynamic changes in protein complex assembly and subunit composition.

Conclusions:

  • Complexome profiling is a robust method for comprehensive interactome analysis.
  • It offers insights into protein complex assembly, supercomplexes, and dynamic processes.
  • The technique is valuable for differential studies in disease models and patient samples.