BBA - Molecular Cell Research (v.1592, #1)
Editorial Board (ii).
Functions of outer membrane receptors in mitochondrial protein import by Toshiya Endo; Daisuke Kohda (3-14).
Most mitochondrial proteins are synthesized in the cytosol as precursor proteins and are imported into mitochondria. The targeting signals for mitochondria are encoded in the presequences or in the mature parts of the precursor proteins, and are decoded by the receptor sites in the translocator complex in the mitochondrial outer membrane. The recently determined NMR structure of the general import receptor Tom20 in a complex with a presequence peptide reveals that, although the amphiphilicity and positive charges of the presequence is essential for the import ability of the presequence, Tom20 recognizes only the amphiphilicity, but not the positive charges. This leads to a new model that different features associated with the mitochondrial targeting sequence of the precursor protein can be recognized by the mitochondrial protein import system in different steps during the import.
Keywords: Mitochondria; Protein import; Receptor; Targeting signal; Presequence;
The mitochondrial import machinery: preprotein-conducting channels with binding sites for presequences by Nikolaus Pfanner; Agnieszka Chacinska (15-24).
Mitochondrial preproteins with amino-terminal presequences must cross two membranes to reach the matrix of the organelle. Both outer and inner membranes contain hydrophilic high-conductance channels that are responsible for selective translocation of preproteins. The channels are embedded in dynamic protein complexes, the TOM complex of the outer membrane and the TIM23 complex of the inner membrane. Both channel-forming proteins, Tom40 and Tim23, carry specific binding sites for presequences, but differ in their pore size and response to a membrane potential. Studies with the TOM machinery show that other subunits of the translocase complex also provide specific binding sites for preproteins, modulate the channel activity and are critical for assembly of the channel.
Keywords: Mitochondrion; Protein sorting; Protein translocation; TOM complex; Channel; Presequence;
Protein import into and across the mitochondrial inner membrane: role of the TIM23 and TIM22 translocons by Robert E Jensen; Cory D Dunn (25-34).
Import of nuclear-encoded mitochondrial proteins requires the action of at least two different import machines, called translocons, in the mitochondrial inner membrane (IM). The TIM23 complex mediates the translocation of proteins into the mitochondria matrix, whereas the TIM22 complex is required for the insertion of polytopic proteins into the IM. While the two translocons are distinct and composed of separate subunits, the essential reactions in each complex are carried out by homologous proteins. In addition, the core components of both the TIM23 and TIM22 translocons have been shown to form aqueous pores in the mitochondrial IM. In this review, we summarize what is known about import of proteins across the mitochondrial IM.
Keywords: Mitochondrial protein import; Translocon; Membrane protein;
Delivery of nascent polypeptides to the mitochondrial surface by Travis Beddoe; Trevor Lithgow (35-39).
Thousands of polypeptides with diverse biochemical properties, some of which are extremely hydrophobic, are targeted from cytoplasmic ribosomes to the surface of mitochondria. Localised synthesis, as well as transient interactions with a wide array of molecular chaperones and other cytoplasmic factors, can promote productive interaction of mitochondrial proteins with the TOM complex to initiate protein import into mitochondria.
Keywords: Ribosome; polypeptide synthesis; mRNA localisation; Molecular chaperone; Protein import; Mitochondrion;
Contact sites between the outer and inner membrane of mitochondria—role in protein transport by Andreas S Reichert; Walter Neupert (41-49).
Many essential functions of mitochondrial metabolism have been studied in the past three decades in considerable depth: oxidative phosphorylation, catabolism of fatty acids, role in nitrogen metabolism, and amino acid metabolism. More recently, other aspects attracted much attention like protein translocation into mitochondria, inheritance of mitochondrial DNA, movement of mitochondria, their fusion and fission, and their involvement in apoptosis, ageing, cancer and other cellular processes. Together with these new views on the function of mitochondria, new ideas on the structure of mitochondria emerged. Here we will discuss the current knowledge about how the membranes of mitochondria are organized and how they interact. Interactions between components of the inner and the outer membrane are necessary for a number of central mitochondrial functions such as the channeling of metabolites, coordinated fusion and fission of mitochondria, and protein transport. Some of these interactions appear stable such as the so-called morphological contact sites; others are quite dynamic. Direct evidence that a certain protein is part of morphologically defined contact sites is lacking. Nevertheless, protein translocase complexes of the outer and the inner membrane exhibit stable interactions between the two membranes when precursor proteins are arrested during import into mitochondria. Finally, we discuss possible roles of cristae junctions, another morphologically defined membrane structure in mitochondria.
Keywords: Mitochondrion; Membrane structure; Protein;
Molecular chaperones as essential mediators of mitochondrial biogenesis by Wolfgang Voos; Karin Röttgers (51-62).
Chaperone proteins have been initially identified by their ability to confer cellular resistance to various stress conditions. However, molecular chaperones participate also in many constitutive cellular processes. Mitochondria contain several members of the major chaperone families that have important functions in maintaining mitochondrial function. The major Hsp70 of the mitochondrial matrix (mtHsp70) is essential for the translocation of cytosolic precursor proteins across the two mitochondrial membranes. MtHsp70 interacts with the preprotein in transit in an ATP-dependent reaction as it emerges from the translocation channel of the inner membrane. Together with two essential partner proteins, Tim44 and Mge1, mtHsp70 forms a membrane-associated import motor complex responsible for vectorial polypeptide movement and unfolding of preprotein domains. Folding of newly imported proteins in the matrix is assisted by the soluble chaperone system formed by mtHsp70 and its partner protein Mdj1. For certain substrate proteins, the protected folding environment that is offered by the large oligomeric Hsp60 complex facilitates further folding reactions. The mitochondrial Hsp70 Ssq1 is involved in the assembly of mitochondrial Fe/S clusters together with another member of the DnaJ family, Jac1. Chaperones of the Clp/Hsp100 family mediate the prevention of aggregation under stress conditions and eventually the degradation of mitochondrial proteins. Together, the chaperones of the mitochondrial matrix form a complex interdependent chaperone network that is essential for most reactions of mitochondrial protein biogenesis.
Keywords: Mitochondrion; Membrane translocation; Degradation; Hsp70; Hsp60; Clp/Hsp100;
Mitochondrial processing peptidases by Oleksandr Gakh; Patrizia Cavadini; Grazia Isaya (63-77).
Three peptidases are responsible for the proteolytic processing of both nuclearly and mitochondrially encoded precursor polypeptides targeted to the various subcompartments of the mitochondria. Mitochondrial processing peptidase (MPP) cleaves the vast majority of mitochondrial proteins, while inner membrane peptidase (IMP) and mitochondrial intermediate peptidase (MIP) process specific subsets of precursor polypeptides. All three enzymes are structurally and functionally conserved across species, and their human homologues begin to be recognized as potential players in mitochondrial disease.
Keywords: Mitochondrion; Protein import; Processing peptidase; Presequence;
Insertion of proteins into the inner membrane of mitochondria: the role of the Oxa1 complex by Rosemary A Stuart (79-87).
The inner mitochondrial membrane harbors a large number of proteins that display a wide range of topological arrangements. The majority of these proteins are encoded in the cell's nucleus, but a few polytopic proteins, all subunits of respiratory chain complexes are encoded by the mitochondrial genome. A number of distinct sorting mechanisms exist to direct these proteins into the mitochondrial inner membrane. One of these pathways involves the export of proteins from the matrix into the inner membrane and is used by both proteins synthesized within the mitochondria, as well as by a subset of nuclear encoded proteins. Prior to embarking on the export pathway, nuclear encoded proteins using this sorting route are initially imported into the mitochondrial matrix from the cytosol, their site of synthesis. Protein export from the matrix into the inner membrane bears similarities to Sec-independent protein export in bacteria and requires the function of the Oxa1 protein. Oxa1 is a component of a general protein insertion site in yeast mitochondrial inner membrane used by both nuclear and mitochondrial DNA encoded proteins. Oxa1 is a member of the conserved Oxa1/YidC/Alb3 protein family found throughout prokaryotes throughout eukaryotes (where it is found in mitochondria and chloroplasts). The evidence to demonstrate that the Oxa1/YidC/Alb3 protein family represents a novel evolutionarily conserved membrane insertion machinery is reviewed here.
Keywords: Oxa1; Yeast; Mitochondria; Protein sorting; Protein insertion; Oxa1/YidC/Alb3;
Membrane protein degradation by AAA proteases in mitochondria by Isabel Arnold; Thomas Langer (89-96).
The inner membrane of mitochondria is one of the protein's richest cellular membranes. The biogenesis of the respiratory chain and ATP-synthase complexes present in this membrane is an intricate process requiring the coordinated function of various membrane-bound proteins including protein translocases and assembly factors. It is therefore not surprising that a distinct quality control system is present in this membrane that selectively removes nonassembled polypeptides and prevents their possibly deleterious accumulation in the membrane. The key components of this system are two AAA proteases, membrane-embedded ATP-dependent proteolytic complexes, which expose their catalytic sites at opposite membrane surfaces. Other components include the prohibitin complex with apparently chaperone-like properties and a regulatory function during proteolysis and a recently identified ATP-binding cassette (ABC) transporter that exports peptides derived from the degradation of membrane proteins from the matrix to the intermembrane space. All of these components are highly conserved during evolution and appear to be ubiquitously present in mitochondria of eukaryotic cells, indicating important cellular functions. This review will summarize our current understanding of this proteolytic system and, in particular, focus on the mechanisms guiding the degradation of membrane proteins by AAA proteases.
Keywords: Proteolysis; AAA protease; Prohibitin; ABC-transporter; Peptide export;
Import and assembly of proteins into mitochondria of mammalian cells by Nicholas J Hoogenraad; Linda A Ward; Michael T Ryan (97-105).
Most of our knowledge regarding the process of protein import into mitochondria has come from research employing fungal systems. This review outlines recent advances in our understanding of this process in mammalian cells. In particular, we focus on the characterisation of cytosolic molecular chaperones that are involved in binding to mitochondrial-targeted preproteins, as well as the identification of both conserved and novel subunits of the import machineries of the outer and inner mitochondrial membranes. We also discuss diseases associated with defects in import and assembly of mitochondrial proteins and what is currently known about the regulation of import in mammals.
Keywords: Import; TOM; TIM; Molecular chaperone; Mitochondrion;