bims-cytox1 2021-08-08 papers

Issue of 2021‒08‒08
three papers selected by
Gavin McStay
Staffordshire University

Genes (Basel). 2021 Jul 01. pii: 1031. [Epub ahead of print]12(7):

Molecular Insights into Mitochondrial Protein Translocation and Human Disease.

Eduardo Ruiz-Pesini, Julio Montoya, David Pacheu-Grau.

In human mitochondria, mtDNA encodes for only 13 proteins, all components of the OXPHOS system. The rest of the mitochondrial components, which make up approximately 99% of its proteome, are encoded in the nuclear genome, synthesized in cytosolic ribosomes and imported into mitochondria. Different import machineries translocate mitochondrial precursors, depending on their nature and the final destination inside the organelle. The proper and coordinated function of these molecular pathways is critical for mitochondrial homeostasis. Here, we will review molecular details about these pathways, which components have been linked to human disease and future perspectives on the field to expand the genetic landscape of mitochondrial diseases.

Keywords: disease; mitochondria; protein import

DOI: https://doi.org/10.3390/genes12071031

Int J Mol Sci. 2021 Jul 21. pii: 7809. [Epub ahead of print]22(15):

Complexome Profiling: Assembly and Remodeling of Protein Complexes.

Ilka Wittig, Pedro Felipe Malacarne.

Many proteins have been found to operate in a complex with various biomolecules such as proteins, nucleic acids, carbohydrates, or lipids. Protein complexes can be transient, stable or dynamic and their association is controlled under variable cellular conditions. Complexome profiling is a recently developed mass spectrometry-based method that combines mild separation techniques, native gel electrophoresis, and density gradient centrifugation with quantitative mass spectrometry to generate inventories of protein assemblies within a cell or subcellular fraction. This review summarizes applications of complexome profiling with respect to assembly ranging from single subunits to large macromolecular complexes, as well as their stability, and remodeling in health and disease.

Keywords: assembly; complexome profiling; data repositories; mass spectrometry; protein complexes; protein–protein interaction; remodeling

DOI: https://doi.org/10.3390/ijms22157809

J Vis Exp. 2021 Jul 19.

Assessment of Submitochondrial Protein Localization in Budding Yeast Saccharomyces cerevisiae.

Fernando Gomes, Helena Turano, Angélica Ramos, Luis E S Netto.

Despite recent advances in the characterization of yeast mitochondrial proteome, the submitochondrial localization of a significant number of proteins remains elusive. Here, we describe a robust and effective method for determining the suborganellar localization of yeast mitochondrial proteins, which is considered a fundamental step during mitochondrial protein function elucidation. This method involves an initial step that consists of obtaining highly pure intact mitochondria. These mitochondrial preparations are then subjected to a subfractionation protocol consisting of hypotonic shock (swelling) and incubation with proteinase K (protease). During swelling, the outer mitochondrial membrane is selectively disrupted, allowing the proteinase K to digest proteins of the intermembrane space compartment. In parallel, to obtain information about the topology of membrane proteins, the mitochondrial preparations are initially sonicated, and then subjected to alkaline extraction with sodium carbonate. Finally, after centrifugation, the pellet and supernatant fractions from these different treatments are analyzed by SDS-PAGE and western blot. The submitochondrial localization as well as the membrane topology of the protein of interest is obtained by comparing its western blot profile with known standards.

DOI: https://doi.org/10.3791/62853