Protein import into mitochondria Mokranjac, D; Neupert, W
Biochemical Society transactions,
11/2005, Letnik:
33, Številka:
Pt 5
Journal Article
Recenzirano
Mitochondria comprise approx. 1000-3000 different proteins, almost all of which must be imported from the cytosol into the organelle. So far, six complex molecular machines, protein translocases, ...were identified that mediate this process. The TIM23 complex is a major translocase in the inner mitochondrial membrane. It uses two energy sources, namely membrane potential and ATP, to facilitate preprotein translocation across the inner membrane and insertion into the inner membrane. Recent research has led to the discovery of a number of new constituents of the TIM23 complex and to the unravelling of the mechanisms of preprotein translocation.
We investigated the effect of substrate binding on the mechanical stability of mouse dihydrofolate reductase using single-molecule force spectroscopy by atomic force microscopy. We find that under ...mechanical forces dihydrofolate reductase unfolds via a metastable intermediate with lifetimes on the millisecond timescale. Based on the measured length increase of ∼22nm we suggest a structure for this intermediate with intact substrate binding sites. In the presence of the substrate analog methotrexate and the cofactor NADPH lifetimes of this intermediate are increased by up to a factor of two. Comparing mechanical and thermodynamic stabilization effects of substrate binding suggests mechanical stability is dominated by local interactions within the protein structure. These experiments demonstrate that protein mechanics can be used to probe the substrate binding status of an enzyme.
PROTEIN IMPORT INTO MITOCHONDRIA Neupert, Walter
Annual review of biochemistry,
01/1997, Letnik:
66, Številka:
1
Journal Article
Recenzirano
Mitochondria import many hundreds of different proteins that are encoded by
nuclear genes. These proteins are targeted to the mitochondria, translocated
through the mitochondrial membranes, and ...sorted to the different mitochondrial
subcompartments. Separate translocases in the mitochondrial outer membrane (TOM
complex) and in the inner membrane (TIM complex) facilitate recognition of
preproteins and transport across the two membranes. Factors in the cytosol
assist in targeting of preproteins. Protein components in the matrix partake in
energetically driving translocation in a reaction that depends on the membrane
potential and matrix-ATP. Molecular chaperones in the matrix exert multiple
functions in translocation, sorting, folding, and assembly of newly imported
proteins.
Celotno besedilo
Dostopno za:
CMK, DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
We examined the effects of ML3000 and several non-steroidal antiinflammatory drugs (NSAIDs) on the synthesis of products of 5-LOX (LTB4, LTC4) and COX-1/2 (TXB2, PGE2) in vitro and ex vivo in order ...to further elucidate the mechanism of action of ML3000.
Using a human whole blood assay the effect of ML3000 on the shunt of arachidonic acid to the lipoxygenase pathway when COX is blocked was studied. ML3000 (0.3, 1, 3, 10, 30 microg/ml) and indomethacin (0.3, 1, 3, 10, 30 microg/ml) concentration-dependently inhibited the synthesis of PGE2 (IC50 = 3.9 and 4.5 microM). In contrast to ML3000, indomethacin produced an increase of LTC4 of up to 155.5% of control. 5-lipoxygenase inhibition was further tested in a basophilic leukemia cell assay using RBL-1 cells. ML3000 (1-10 microM) inhibited the synthesis of LTB4 in a concentration related manner (IC50: 3.6 microM). In carrageenan induced rat paw edema, ML3000 and indomethacin completely blocked the formation of PGE2 in the inflamed tissue. The LTB4 production in the inflamed paw was reduced to basal levels by ML3000 (10 +/- 1.4 pg/paw saline control and 7.5 +/- 1.3-5.9 +/- 3.2 pg/paw ML3000), whereas LTB4 levels remained markedly elevated as compared to saline control by indomethacin (30.7 pg/paw). 5-LOX inhibition in the inflamed rat colon was investigated by measuring LTB4 synthesis. MK-886 and ML3000 at 10 mg/kg p.o. reduced LTB4 production to 29.8 +/- 4.9 and 30.1 +/- 2.8 pg/mg tissue as compared to control (54.2 +/- 7.4 mg/kg tissue). LTB4 levels in the rat stomach were comparable to control (2.5 +/- 0.4 pg/mg protein) after oral administration of ML3000 (10, 30, 100 mg/kg), whereas oral treatment with indomethacin (0.3, 1, 3 mg/kg) or diclofenac (1, 3 mg/kg) increased LTB4 up to 9.2 +/- 2.3 or 8.9 +/- 1.6 pg/mg protein. This effect was significant at 1 mg/kg diclofenac and 0.3 mg/kg indomethacin.
These results provide further evidence, that ML3000 inhibits 5-LOX as well as COX-1 and COX-2 in vitro and in animal experiments. The favourable gastrointestinal (GI) tolerability of the compound is believed to be linked to the mechanism of combined 5-LOX and COX-1/2 inhibition of ML3000.
Import of nuclear-encoded precursor proteins into mitochondria and their subsequent sorting into mitochondrial subcompartments is mediated by translocase enzymes in the mitochondrial outer and inner ...membranes. Precursor proteins carrying amino-terminal targeting signals are translocated into the matrix by the integral inner membrane proteins Tim23 and Tim17 in cooperation with Tim44 and mitochondrial Hsp70 (refs 4-7). We describe here the discovery of a new pathway for the transport of members of the mitochondrial carrier family and other inner membrane proteins that contain internal targeting signals. Two related proteins in the intermembrane space, Tim10/Mrs11 (ref. 8) and Tim12/Mrs5 (ref. 9), interact sequentially with these precursors and facilitate their translocation across the outer membrane, irrespective of the membrane potential. Tim10 and Tim12 are found in a complex with Tim22, which takes over the precursor and mediates its membrane-potential-dependent insertion into the inner membrane. This interaction of Tim10 and Tim12 with the precursors depends on the presence of divalent metal ions. Both proteins contain a zinc-finger-like motif with four cysteines and bind equimolar amounts of zinc ions.
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Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Protein Import Into Mitochondria Paschen, Stefan A.; Neupert, Walter
IUBMB life,
1 September 2001, Letnik:
52, Številka:
3‐5
Journal Article
Recenzirano
Odprti dostop
Most mitochondrial proteins are encoded by the nuclear genome and thus have to be imported into mitochondria from the cytosol. Protein translocation across and into the mitochondrial membranes is a ...multistep process facilitated by the coordinated action of at least four specialized translocation systems in the outer and inner membranes of mitochondria. The outer membrane contains one general translocase, the TOM complex, whereas three distinct translocases are located in the inner membrane, which facilitates translocation of different classes of preproteins. The TIM23 complex mediates import of matrix‐targeted preproteins with N ‐terminal presequences, whereas hydrophobic preproteins with internal targeting signals are inserted into the inner membrane via the TIM22 complex. The OXA translocase mediates the insertion of preproteins from the matrix space into the inner membrane. This review focuses on the structural organization and function of the import machinery of the model organisms of Saccharomyces cerevisiae and Neurospora crassa . In addition, the molecular basis of a new human mitochondrial disorder is discussed, the Mohr‐Tranebjaerg syndrome. This is the first known disease, which is caused by an impaired mitochondrial protein import machinery leading to progressive neurodegeneration.
The Extreme Ultraviolet Imaging Telescope (EIT) on board the SOHO spacecraft has been operational since 2 January 1996. EIT observes the Sun over a 45 x 45 arc min field of view in four emission line ...groups: Feix, x, Fexii, Fexv, and Heii. A post-launch determination of the instrument flatfield, the instrument scattering function, and the instrument aging were necessary for the reduction and analysis of the data. The observed structures and their evolution in each of the four EUV bandpasses are characteristic of the peak emission temperature of the line(s) chosen for that bandpass. Reports on the initial results of a variety of analysis projects demonstrate the range of investigations now underway: EIT provides new observations of the corona in the temperature range of 1 to 2 MK. Temperature studies of the large-scale coronal features extend previous coronagraph work with low-noise temperature maps. Temperatures of radial, extended, plume-like structures in both the polar coronal hole and in a low latitude decaying active region were found to be cooler than the surrounding material. Active region loops were investigated in detail and found to be isothermal for the low loops but hottest at the loop tops for the large loops.Variability of solar EUV structures, as observed in the EIT time sequences, is pervasive and leads to a re-evaluation of the meaning of the term 'quiet Sun'. Intensity fluctuations in a high cadence sequence of coronal and chromospheric images correspond to a Kolmogorov turbulence spectrum. This can be interpreted in terms of a mixed stochastic or periodic driving of the transition region and the base of the corona. No signature of the photospheric and chromospheric waves is found in spatially averaged power spectra, indicating that these waves do not propagate to the upper atmosphere or are channeled through narrow local magnetic structures covering a small fraction of the solar surface. Polar coronal hole observing campaigns have identified an outflow process with the discovery of transient Fexii jets. Coronal mass ejection observing campaigns have identified the beginning of a CME in an Fexii sequence with a near simultaneous filament eruption (seen in absorption), formation of a coronal void and the initiation of a bright outward-moving shell as well as the coronal manifestation of a 'Moreton wave'.
The protein import motor of mitochondria Neupert, Walter; Brunner, Michael
Nature reviews. Molecular cell biology,
08/2002, Letnik:
3, Številka:
8
Journal Article
Recenzirano
Proteins that are destined for the matrix of mitochondria are transported into this organelle by two translocases: the TOM complex, which transports proteins across the outer mitochondrial membrane; ...and the TIM23 complex, which gets them through the inner mitochondrial membrane. Two models have been proposed to explain how this protein-import machinery works -- a targeted Brownian ratchet, in which random motion is translated into vectorial motion, or a 'power stroke', which is exerted by a component of the import machinery. Here, we review the data for and against each model.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
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