scholarly journals The receptor subunit Tom20 is dynamically associated with the TOM complex in mitochondria of human cells

2021 ◽  
pp. mbc.E21-01-0042
Author(s):  
Maniraj Bhagawati ◽  
Tasnim Arroum ◽  
Niklas Webeling ◽  
Ayelén González Montoro ◽  
Henning D. Mootz ◽  
...  
Keyword(s):  
Single Particle Tracking ◽  
Human Cells ◽  
Receptor Subunit ◽  
Core Complex ◽  
Tom Complex ◽  
System A ◽  
Precursor Proteins ◽  
The Mean ◽  
Reduced Mobility ◽  
Peripheral Receptor

The outer membrane translocase (TOM) is the import channel for nuclear-encoded mitochondrial proteins. The general import pore contains Tom40, Tom22, Tom5, Tom6 and Tom7. Precursor proteins are bound by the peripheral receptor proteins Tom20, Tom22 and Tom70 before being imported by the TOM complex. Here we investigated the association of the receptor Tom20 with the TOM complex. Tom20 was found in the TOM complex, but not in a smaller subcomplex. In addition, a subcomplex was found without Tom40 and Tom7 but with Tom20. Using single particle tracking of labeled Tom20 in overexpressing human cells, we show that Tom20 has, on average, higher lateral mobility in the membrane than Tom7/TOM. After ligation of Tom20 with the TOM complex by post-tranlational protein trans-splicing using the trackless, ultra-fast cleaved Gp41-1 integrin system, a significant decrease in the mean diffusion coefficient of Tom20 was observed in the resulting Tom20-Tom7 fusion protein. Exposure of Tom20 to high substrate loading also resulted in reduced mobility. Taken together, our data show that the receptor subunit Tom20 interacts dynamically with the TOM core complex. We suggest that the TOM complex containing Tom20 is the active import pore and that Tom20 is associated when substrate is available.

scholarly journals Reconstituted TOM Core Complex and Tim9/Tim10 Complex of Mitochondria Are Sufficient for Translocation of the ADP/ATP Carrier across Membranes

2004 ◽  
Vol 15 (3) ◽  
pp. 1445-1458 ◽  
Author(s):  
Andreja Vasiljev ◽  
Uwe Ahting ◽  
Frank E. Nargang ◽  
Nancy E. Go ◽  
Shukry J. Habib ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Lipid Vesicles ◽  
Intermembrane Space ◽  
Core Complex ◽  
Tom Complex ◽  
Precursor Proteins ◽  
Solute Carrier ◽  
Membrane Peptide ◽  
Membrane Spanning ◽  
Substrate Proteins

Precursor proteins of the solute carrier family and of channel forming Tim components are imported into mitochondria in two main steps. First, they are translocated through the TOM complex in the outer membrane, a process assisted by the Tim9/Tim10 complex. They are passed on to the TIM22 complex, which facilitates their insertion into the inner membrane. In the present study, we have analyzed the function of the Tim9/Tim10 complex in the translocation of substrates across the outer membrane of mitochondria. The purified TOM core complex was reconstituted into lipid vesicles in which purified Tim9/Tim10 complex was entrapped. The precursor of the ADP/ATP carrier (AAC) was found to be translocated across the membrane of such lipid vesicles. Thus, these components are sufficient for translocation of AAC precursor across the outer membrane. Peptide libraries covering various substrate proteins were used to identify segments that are bound by Tim9/Tim10 complex upon translocation through the TOM complex. The patterns of binding sites on the substrate proteins suggest a mechanism by which portions of membrane-spanning segments together with flanking hydrophilic segments are recognized and bound by the Tim9/Tim10 complex as they emerge from the TOM complex into the intermembrane space.

scholarly journals Role of Phosphatidylethanolamine in the Biogenesis of Mitochondrial Outer Membrane Proteins

2013 ◽  
Vol 288 (23) ◽  
pp. 16451-16459 ◽  
Author(s):  
Thomas Becker ◽  
Susanne E. Horvath ◽  
Lena Böttinger ◽  
Natalia Gebert ◽  
Günther Daum ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Outer Membrane ◽  
Outer Membrane Proteins ◽  
Proper Function ◽  
Mitochondrial Outer Membrane ◽  
Tom Complex ◽  
Precursor Proteins ◽  
Helical Proteins ◽  
The Stability

The mitochondrial outer membrane contains proteinaceous machineries for the import and assembly of proteins, including TOM (translocase of the outer membrane) and SAM (sorting and assembly machinery). It has been shown that the dimeric phospholipid cardiolipin is required for the stability of TOM and SAM complexes and thus for the efficient import and assembly of β-barrel proteins and some α-helical proteins of the outer membrane. Here, we report that mitochondria deficient in phosphatidylethanolamine (PE), the second non-bilayer-forming phospholipid, are impaired in the biogenesis of β-barrel proteins, but not of α-helical outer membrane proteins. The stability of TOM and SAM complexes is not disturbed by the lack of PE. By dissecting the import steps of β-barrel proteins, we show that an early import stage involving translocation through the TOM complex is affected. In PE-depleted mitochondria, the TOM complex binds precursor proteins with reduced efficiency. We conclude that PE is required for the proper function of the TOM complex.

scholarly journals A discrete pathway for the transfer of intermembrane space proteins across the outer membrane of mitochondria

2014 ◽  
Vol 25 (25) ◽  
pp. 3999-4009 ◽  
Author(s):  
Agnieszka Gornicka ◽  
Piotr Bragoszewski ◽  
Piotr Chroscicki ◽  
Lena-Sophie Wenz ◽  
Christian Schulz ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Mitochondrial Membrane ◽  
Alternative Pathway ◽  
Intermembrane Space ◽  
Outer Mitochondrial Membrane ◽  
Membrane Translocation ◽  
Tom Complex ◽  
Precursor Proteins ◽  
Core Components

Mitochondrial proteins are synthesized on cytosolic ribosomes and imported into mitochondria with the help of protein translocases. For the majority of precursor proteins, the role of the translocase of the outer membrane (TOM) and mechanisms of their transport across the outer mitochondrial membrane are well recognized. However, little is known about the mode of membrane translocation for proteins that are targeted to the intermembrane space via the redox-driven mitochondrial intermembrane space import and assembly (MIA) pathway. On the basis of the results obtained from an in organello competition import assay, we hypothesized that MIA-dependent precursor proteins use an alternative pathway to cross the outer mitochondrial membrane. Here we demonstrate that this alternative pathway involves the protein channel formed by Tom40. We sought a translocation intermediate by expressing tagged versions of MIA-dependent proteins in vivo. We identified a transient interaction between our model substrates and Tom40. Of interest, outer membrane translocation did not directly involve other core components of the TOM complex, including Tom22. Thus MIA-dependent proteins take another route across the outer mitochondrial membrane that involves Tom40 in a form that is different from the canonical TOM complex.

Multiple channel queues in heavy traffic. I

1970 ◽  
Vol 2 (01) ◽  
pp. 150-177 ◽  
Author(s):  
Donald L. Iglehart ◽  
Ward Whitt
Keyword(s):  
Heavy Traffic ◽  
Arrival Rate ◽  
Standard System ◽  
Interarrival Time ◽  
Service Rate ◽  
Service Channel ◽  
Multiple Channel ◽  
System A ◽  
The Mean ◽  
Service Channels

The queueing systems considered in this paper consist of r independent arrival channels and s independent service channels, where as usual the arrival and service channels are independent. Arriving customers form a single queue and are served in the order of their arrival without defections. We shall treat two distinct modes of operation for the service channels. In the standard system a waiting customer is assigned to the first available service channel and the servers (servers ≡ service channels) are shut off when they are idle. Thus the classical GI/G/s system is a special case of our standard system. In the modified system a waiting customer is assigned to the service channel that can complete his service first and the servers are not shut off when they are idle. While the modified system is of some interest in its own right, we introduce it primarily as an analytical tool. Let λ i denote the arrival rate (reciprocal of the mean interarrival time) in the ith arrival channel and μ j the service rate (reciprocal of the mean service time) in the jth service channel. Then is the total arrival rate to the system and is the maximum service rate of the system. As a measure of congestion we define the traffic intensity ρ = λ/μ.

scholarly journals Functions of the Small Proteins in the TOM Complex of Neurospora crasssa

2005 ◽  
Vol 16 (9) ◽  
pp. 4172-4182 ◽  
Author(s):  
E. Laura Sherman ◽  
Nancy E. Go ◽  
Frank E. Nargang
Keyword(s):  
Mitochondrial Membrane ◽  
Minor Role ◽  
Complex Stability ◽  
Outer Mitochondrial Membrane ◽  
The Core ◽  
Tom Complex ◽  
Membrane Complex ◽  
Small Proteins ◽  
Precursor Proteins ◽  
A Minor

The TOM (translocase of the outer mitochondrial membrane) complex of the outer mitochondrial membrane is required for the import of proteins into the organelle. The core TOM complex contains five proteins, including three small components Tom7, Tom6, and Tom5. We have created single and double mutants of all combinations of the three small Tom proteins of Neurospora crassa. Analysis of the mutants revealed that Tom6 plays a major role in TOM complex stability, whereas Tom7 has a lesser role. Mutants lacking both Tom6 and Tom7 have an extremely labile TOM complex and are the only class of mutant to exhibit an altered growth phenotype. Although single mutants lacking N. crassa Tom5 have no apparent TOM complex abnormalities, studies of double mutants lacking Tom5 suggest that it also has a minor role in maintaining TOM complex stability. Our inability to isolate triple mutants supports the idea that the three proteins have overlapping functions. Mitochondria lacking either Tom6 or Tom7 are differentially affected in their ability to import different precursor proteins into the organelle, suggesting that they may play roles in the sorting of proteins to different mitochondrial subcompartments. Newly imported Tom40 was readily assembled into the TOM complex in mitochondria lacking any of the small Tom proteins.

scholarly journals Characterization of the translocon of the outer envelope of chloroplasts

2003 ◽  
Vol 160 (4) ◽  
pp. 541-551 ◽  
Author(s):  
Enrico Schleiff ◽  
Jürgen Soll ◽  
Michael Küchler ◽  
Werner Kühlbrandt ◽  
Roswitha Harrer
Keyword(s):  
Three Dimensional ◽  
Dependent Manner ◽  
Interior Structure ◽  
Two Dimensional ◽  
Outer Envelope ◽  
Core Complex ◽  
The Core ◽  
Precursor Proteins ◽  
Stable Core

The protein translocon of the outer envelope of chloroplasts (Toc) consists of the core subunits Toc159, Toc75, and Toc34. To investigate the molecular structure, the core complex was purified. This core complex has an apparent molecular mass of ∼500 kD and a molecular stoichiometry of 1:4:4–5 between Toc159, Toc75, and Toc34. The isolated translocon recognizes both transit sequences and precursor proteins in a GTP-dependent manner, suggesting its functional integrity. The complex is embedded by the lipids phosphatidylcholine and digalactosyldiacylglyceride. Two-dimensional structural analysis by EM revealed roughly circular particles consistent with the formation of a stable core complex. The particles show a diameter of ∼130 Å with a solid ring and a less dense interior structure. A three-dimensional map obtained by random conical tilt reconstruction of electron micrographs suggests that a “finger”-like central region separates four curved translocation channels within one complex.

scholarly journals A Flow-Calibrated Method to Project Groundwater Infiltration into Coastal Sewers Affected by Sea Level Rise

Water ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 1934
Author(s):  
Adrienne Fung ◽  
Roger Babcock
Keyword(s):  
Sea Level Rise ◽  
Sea Level ◽  
Monitoring Data ◽  
Groundwater Levels ◽  
Flow Monitoring ◽  
System A ◽  
Coastal Cities ◽  
The Mean ◽  
Groundwater Infiltration

Collection systems in coastal cities are often below the groundwater table, leading to groundwater infiltration (GWI) through defects such as cracks and poor lateral connections. Climate-change-induced sea level rise (SLR) will raise groundwater levels, increasing the head and thus the inflow. A method has been developed to predict GWI when groundwater levels change using calibration with sewershed flow monitoring data. The calibration results in a parameter that characterizes the porosity of the collection system. A case study is presented for a coastal city with reliable flow monitoring data for eight days that resulted in a large range of effective defect sizes (minimum 0.0044 to maximum 0.338 radians), however, the range of predicted future GWI in currently submerged pipes varied by only 12% from the mean. The mean effective defect predicts 70 to 200% increases in GWI due to SLR of 0.3 to 0.9 m (1 to 3 ft), respectively, for currently submerged pipes. Predicted additional GWI for pipes that will become submerged due to SLR will increase GWI to values that approach or exceed the current average dry weather flow. This methodology can be used for planning of infrastructure improvements to enhance resiliency in coastal communities.

ABOUT THE MEAN KING'S PROBLEM AND DISCRETE WIGNER DISTRIBUTIONS

2006 ◽  
Vol 20 (11n13) ◽  
pp. 1742-1760 ◽  
Author(s):  
THOMAS DURT
Keyword(s):  
Weyl Function ◽  
Galois Field ◽  
Bell States ◽  
Quadratic Extensions ◽  
System A ◽  
Wigner Distributions ◽  
Particular Solution ◽  
The Mean ◽  
One To One ◽  
Adjoint Operators

When the state of a quantum system belongs to a N-dimensional Hilbert space, with N the power of a prime number, it is possible to associate to the system a finite field (Galois field) with N elements. In this paper, we introduce generalized Bell states that can be intrinsically expressed in terms of the field operations. These Bell states are in one to one correspondence with the N2elements of the generalised Pauli group or Heisenberg-Weyl group. This group consists of discrete displacement operators and provides a discrete realisation of the Weyl function. Thanks to the properties of generalised Bell states and of quadratic extensions of finite fields, we derive a particular solution for the Mean King's problem. This solution is in turn shown to be in one to one correspondence with a set of N2self-adjoint operators that provides a discrete realisation of the Wigner quasi-distribution.

Analysis of an Artificial Ventricle and Mock Circulatory System

1977 ◽  
Vol 99 (4) ◽  
pp. 184-188 ◽  
Author(s):  
K. M. High ◽  
J. A. Brighton ◽  
A. D. Brickman ◽  
W. S. Pierce
Keyword(s):  
Artificial Heart ◽  
Circulatory System ◽  
Experimental Tests ◽  
Mean Flow ◽  
Actual System ◽  
Mock Circulatory System ◽  
System A ◽  
The Mean ◽  
Artificial Ventricle ◽  
Good Agreement

A mathematical model is developed for calculating the pressures and flows in an artificial heart, its pneumatic drive unit, and a mock circulatory system. The system is divided into convenient subsystems to facilitate the analysis, and each subsystem is then analyzed separately. The set of independent equations developed is solved on a computer and corresponding experimental tests are made on the actual system. A comparison of the experimental and computer results shows good agreement for the mean flow rate through the pump and also for several instantaneous pressures and flow rates in the system.

scholarly journals Structural Requirements of Tom40 for Assembly into Preexisting TOM Complexes of Mitochondria

2001 ◽  
Vol 12 (5) ◽  
pp. 1189-1198 ◽  
Author(s):  
Doron Rapaport ◽  
Rebecca D. Taylor ◽  
Michael Käser ◽  
Thomas Langer ◽  
Walter Neupert ◽  
...  
Keyword(s):  
Intermediate Stage ◽  
In Vivo Studies ◽  
Mitochondrial Outer Membrane ◽  
Amino Acid Residues ◽  
Wild Type ◽  
Tom Complex ◽  
Precursor Proteins ◽  
Blue Native

Tom40 is the major subunit of the translocase of the outer mitochondrial membrane (the TOM complex). To study the assembly pathway of Tom40, we have followed the integration of the protein into the TOM complex in vitro and in vivo using wild-type and altered versions of the Neurospora crassa Tom40 protein. Upon import into isolated mitochondria, Tom40 precursor proteins lacking the first 20 or the first 40 amino acid residues were assembled as the wild-type protein. In contrast, a Tom40 precursor lacking residues 41 to 60, which contains a highly conserved region of the protein, was arrested at an intermediate stage of assembly. We constructed mutant versions of Tom40 affecting this region and transformed the genes into a sheltered heterokaryon containing a tom40 null nucleus. Homokaryotic strains expressing the mutant Tom40 proteins had growth rate defects and were deficient in their ability to form conidia. Analysis of the TOM complex in these strains by blue native gel electrophoresis revealed alterations in electrophoretic mobility and a tendency to lose Tom40 subunits from the complex. Thus, both in vitro and in vivo studies implicate residues 41 to 60 as containing a sequence required for proper assembly/stability of Tom40 into the TOM complex. Finally, we found that TOM complexes in the mitochondrial outer membrane were capable of exchanging subunits in vitro. A model is proposed for the integration of Tom40 subunits into the TOM complex.

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