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Proteomics01:33

Proteomics

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A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
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Mitochondrial Protein Sorting01:39

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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.
Most of these mitochondrial proteins are encoded by the nucleus and imported to the mitochondria as unfolded or loosely folded precursors. Mitochondrial precursors...
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Translocation of Proteins into the Mitochondria01:19

Translocation of Proteins into the Mitochondria

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Mitochondrial precursors are translocated to the internal subcompartments via independent mechanisms involving distinct protein machineries called translocases.
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Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
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Porin Insertion in the Outer Mitochondrial Membrane01:12

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Porins are beta-barrel proteins translocated to the mitochondrial outer membrane through the TOM complex into the intermembrane space. Porin precursors bind TIM chaperones within the intermembrane space and are guided to the Sorting and Assembly Machinery complex or SAM complex on the outer mitochondrial membrane.
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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.
Most of the mitochondrial...
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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
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Preparation of Mitochondrial Enriched Fractions for Metabolic Analysis in Drosophila
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Definición de las funciones de las proteínas mitocondriales a través de un perfil multiómico profundo

Jarred W Rensvold1,2, Evgenia Shishkova3,4, Yuriy Sverchkov5

  • 1Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO, USA.

Nature
|May 25, 2022
PubMed
Resumen
Este resumen es generado por máquina.

Los investigadores perfilaron más de 200 nocauts de proteínas mitocondriales para mapear las respuestas celulares. Este estudio identificó nuevas funciones genéticas y una firma para diagnosticar trastornos mitocondriales.

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Área de la Ciencia:

  • Biología celular
  • La genética
  • La bioquímica

Sus antecedentes:

  • Las mitocondrias son vitales para la energía celular y el metabolismo.
  • La disfunción mitocondrial causa más de 150 trastornos, sin embargo, muchas proteínas y causas genéticas siguen siendo desconocidas.
  • Comprender la función de las proteínas mitocondriales es crucial para diagnosticar y tratar estas enfermedades.

Objetivo del estudio:

  • Para crear un mapa funcional completo de las proteínas mitocondriales humanas.
  • Para investigar las respuestas celulares a las perturbaciones mitocondriales.
  • Para identificar nuevos genes asociados con trastornos mitocondriales.

Principales métodos:

  • Utilizó la edición de genes CRISPR-Cas9 para crear más de 200 líneas knockout de células HAP1 dirigidas a proteínas mitocondriales.
  • Se realizaron análisis multiómicos basados en la espectrometría de masas para medir las respuestas celulares.
  • Se recogieron aproximadamente 8,3 millones de mediciones de biomoléculas.

Principales resultados:

  • Se identificó PIGY upstream open reading frame (PYURF) como un chaperón de la metiltransferasa involucrado en el ensamblaje del complejo I y la biosíntesis de la coenzima Q, vinculado a un nuevo trastorno mitocondrial.
  • SLC30A9 vinculado a los ribosomas mitocondriales y la integridad de la fosforilación oxidativa (OxPhos).
  • Establecido RAB5IF como un segundo gen que causa displasia cerebrofaciotorácica.
  • Generó un conjunto de datos completo de las funciones de las proteínas mitocondriales y las respuestas celulares.
  • Descubrió una firma celular que indica una disfunción mitocondrial.

Conclusiones:

  • El estudio proporciona un compendio funcional de proteínas mitocondriales humanas, ayudando en la caracterización de proteínas huérfanas.
  • Identificó nuevas funciones genéticas y asociaciones de enfermedades, avanzando en la comprensión de los trastornos mitocondriales.
  • El recurso MITOMICS.app facilita la exploración de las funciones de las proteínas mitocondriales y los mecanismos de las enfermedades.
  • La firma celular identificada ayuda en el diagnóstico genético de las enfermedades mitocondriales.