Experimental approaches to the study of the biogenesis of mammalian mitochondrial proteins

1986 ◽  
Vol 64 (11) ◽  
pp. 1108-1114 ◽  
Author(s):  
Karl B. Freeman ◽  
Randall W. Yatscoff ◽  
Robert G. Ridley
Keyword(s):  
Mitochondrial Proteins ◽  
Specific Inhibition ◽  
Mitochondrial Translation ◽  
Isolated Mitochondria ◽  
Approaches To Study ◽  
Experimental Approaches ◽  
Specific Inhibitors ◽  
Cytosolic Ribosomes

Mitochondrial proteins are synthesized in mitochondria and on cytosolic ribosomes. Several approaches used to establish the site of synthesis and the identity of mitochondrially synthesized proteins are described. These include the specific inhibition of mitochondrial translation by inhibitors or mutation and the specific elimination of cytosolic translation either by using isolated mitochondria or specific inhibitors. Experimental approaches to study the import of proteins into mitochondria are also discussed.

scholarly journals Advanced experimental approaches to marine water-column biogeochemical processes

2017 ◽  
Vol 75 (1) ◽  
pp. 30-42 ◽  
Author(s):  
Louis Legendre ◽  
Richard B Rivkin ◽  
Nianzhi Jiao
Keyword(s):  
Water Column ◽  
Numerical Models ◽  
Free Water ◽  
Sea Surface ◽  
Biogeochemical Processes ◽  
In Situ Experiments ◽  
Technological Developments ◽  
Approaches To Study ◽  
Experimental Approaches

Abstract This “Food for Thought” article examines the potential uses of several novel scientific and technological developments, which are currently available or being developed, to significantly advance or supplement existing experimental approaches to study water-column biogeochemical processes (WCB-processes). After examining the complementary roles of observation, experiments and numerical models to study WCB-processes, we focus on the main experimental approaches of free-water in situ experiments, and at-sea and on-land meso- and macrocosms. We identify some of the incompletely resolved aspects of marine WCB-processes, and explore advanced experimental approaches that could be used to reduce their uncertainties. We examine three such approaches: free-water experiments of lengthened duration using bioArgo floats and gliders, at-sea mesocosms deployed several 100s m below the sea-surface using new biogeochemical sensors, and 50 m-tall on-land macrocosms. These approaches could lead to significant progress in concepts related to marine WCB-processes.

scholarly journals Novel in vitro Experimental Approaches to Study Myelination and Remyelination in the Central Nervous System

2021 ◽  
Vol 15 ◽  
Author(s):  
Davide Marangon ◽  
Nicolò Caporale ◽  
Marta Boccazzi ◽  
Maria P. Abbracchio ◽  
Giuseppe Testa ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Molecular Mechanisms ◽  
Disease Modeling ◽  
Brain Physiology ◽  
Pathological Conditions ◽  
Approaches To Study ◽  
The Central Nervous System ◽  
Experimental Approaches

Myelin is the lipidic insulating structure enwrapping axons and allowing fast saltatory nerve conduction. In the central nervous system, myelin sheath is the result of the complex packaging of multilamellar extensions of oligodendrocyte (OL) membranes. Before reaching myelinating capabilities, OLs undergo a very precise program of differentiation and maturation that starts from OL precursor cells (OPCs). In the last 20 years, the biology of OPCs and their behavior under pathological conditions have been studied through several experimental models. When co-cultured with neurons, OPCs undergo terminal maturation and produce myelin tracts around axons, allowing to investigate myelination in response to exogenous stimuli in a very simple in vitro system. On the other hand, in vivo models more closely reproducing some of the features of human pathophysiology enabled to assess the consequences of demyelination and the molecular mechanisms of remyelination, and they are often used to validate the effect of pharmacological agents. However, they are very complex, and not suitable for large scale drug discovery screening. Recent advances in cell reprogramming, biophysics and bioengineering have allowed impressive improvements in the methodological approaches to study brain physiology and myelination. Rat and mouse OPCs can be replaced by human OPCs obtained by induced pluripotent stem cells (iPSCs) derived from healthy or diseased individuals, thus offering unprecedented possibilities for personalized disease modeling and treatment. OPCs and neural cells can be also artificially assembled, using 3D-printed culture chambers and biomaterial scaffolds, which allow modeling cell-to-cell interactions in a highly controlled manner. Interestingly, scaffold stiffness can be adopted to reproduce the mechanosensory properties assumed by tissues in physiological or pathological conditions. Moreover, the recent development of iPSC-derived 3D brain cultures, called organoids, has made it possible to study key aspects of embryonic brain development, such as neuronal differentiation, maturation and network formation in temporal dynamics that are inaccessible to traditional in vitro cultures. Despite the huge potential of organoids, their application to myelination studies is still in its infancy. In this review, we shall summarize the novel most relevant experimental approaches and their implications for the identification of remyelinating agents for human diseases such as multiple sclerosis.

Interference with Murein Turnover Has No Effect on Growth but Reduces β-Lactamase Induction in Escherichia coli

1999 ◽  
Vol 181 (23) ◽  
pp. 7192-7198 ◽  
Author(s):  
Angelika R. Kraft ◽  
Julia Prabhu ◽  
Astrid Ursinus ◽  
Joachim-Volker Höltje
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Enterobacter Cloacae ◽  
Low Molecular Weight ◽  
Specific Inhibition ◽  
Triple Mutant ◽  
Induced Expression ◽  
Lytic Transglycosylases ◽  
Reduced Rate ◽  
Specific Inhibitors

ABSTRACT Physiological studies of a mutant of Escherichia colilacking the three lytic transglycosylases Slt70, MltA, and MltB revealed that interference with murein turnover can prevent AmpC β-lactamase induction. The triple mutant, although growing normally, shows a dramatically reduced rate of murein turnover. Despite the reduction in the formation of low-molecular-weight murein turnover products, neither the rate of murein synthesis nor the amount of murein per cell was increased. This might be explained by assuming that during growth in the absence of the major lytic transglycosylases native murein strands are excised by the action of endopeptidases and directly reused without further breakdown to muropeptides. The reduced rate of murein turnover could be correlated with lowered cefoxitin-induced expression of β-lactamase, present on a plasmid carrying theampC and ampR genes from Enterobacter cloacae. Overproduction of MltB stimulated β-lactamase induction, whereas specific inhibition of Slt70 by bulgecin repressedampC expression. Thus, specific inhibitors of lytic transglycosylases can increase the potency of penicillins and cephalosporins against bacteria inducing AmpC-like β-lactamases.

Towards astrobiological experimental approaches to study planetary UV surface environments

2018 ◽  
Vol 14 (S345) ◽  
pp. 222-226
Author(s):  
Ximena C. Abrevaya ◽  
Martin Leitzinger ◽  
Oscar J. Oppezzo ◽  
Petra Odert ◽  
G. Juan M. Luna ◽  
...  
Keyword(s):  
Ultraviolet Radiation ◽  
Laboratory Experiments ◽  
Lethal Dose ◽  
Action Spectrum ◽  
Interdisciplinary Approach ◽  
Accurate Estimation ◽  
Atmospheric Attenuation ◽  
Approaches To Study ◽  
Experimental Approaches ◽  
The Impact

AbstractThe stellar ultraviolet radiation (UVR) has been studied in the last decade and has been found to be an important factor to determine the habitability of planetary surfaces. It is known that UVR can be a constraint for life. However, most of the studies of UVR and habitability have missed some fundamental aspects: i) Accurate estimation of the planetary atmospheric attenuation, ii) The biological inferences used to represent the impact of the stellar UVR on life are theoretical and based on the action spectrum (for DNA or microorganisms) or considering parameters as the “lethal dose” obtained from non-astrobiological experiments. Therefore, the conclusions reached by previous studies about the UVR habitability of planetary bodies may be inaccurate. In this work, we propose how to address these studies in a more accurate way through an interdisciplinary approach that combines astrophysics, microbiology, and photobiology and by the use of specially designed laboratory experiments.

Insulin stimulates mitochondrial protein synthesis and respiration in isolated perfused rat heart.

1990 ◽  
Vol 259 (3) ◽  
pp. E413
Author(s):  
E E McKee ◽  
B L Grier
Keyword(s):  
Protein Synthesis ◽  
Mitochondrial Protein ◽  
Control Level ◽  
Respiratory Control ◽  
Mitochondrial Proteins ◽  
Mitochondrial Protein Synthesis ◽  
Mitochondrial Translation ◽  
Rat Hearts ◽  
Perfused Rat Hearts ◽  
Perfused Hearts

The rates of synthesis of mitochondrial proteins by both the cytoplasmic and mitochondrial protein synthetic systems, as well as parameters of respiration, were measured and compared in mitochondria isolated from fresh, control perfused, and insulin-perfused rat hearts. The respiratory control ratio (RCR) in mitochondria from fresh hearts was 8.1 +/- 0.4 and decreased to 6.0 +/- 0.2 (P less than 0.001 vs. fresh) in mitochondria from control perfused hearts and to 6.7 +/- 0.2 (P less than 0.005 vs. fresh and P less than 0.02 vs. control perfused) for mitochondria from hearts perfused in the presence of insulin. A positive correlation between the RCR and the rate of mitochondrial translation was demonstrated in mitochondria from fresh hearts. In mitochondria isolated from control perfused hearts, the rate of protein synthesis decreased to 84 +/- 3% of the fresh rate after 30 min of perfusion and fell further to 64 +/- 3% after 3 h of perfusion. The inclusion of insulin in the perfusion buffer stimulated mitochondrial protein synthesis 1.2-fold by 1 h (P less than 0.005) and 1.34-fold by 3 h of perfusion (P less than 0.001). The addition of insulin to 1-h control perfused hearts shifted the rate of mitochondrial protein synthesis from the control level to the insulin-perfused level within 30 min of additional perfusion, whereas 1 h was required to shift the RCR values of these mitochondria from control levels to insulin-perfused levels. Thus, whereas RCR was a useful predictor of mitochondrial translation rates, it did not account for the effects of insulin on mitochondrial translation.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals The synthesis of murine ferrochelatase in vitro and in vivo

1988 ◽  
Vol 254 (3) ◽  
pp. 799-803 ◽  
Author(s):  
S R Karr ◽  
H A Dailey
Keyword(s):  
Membrane Protein ◽  
Biosynthetic Pathway ◽  
Integral Membrane Protein ◽  
Mitochondrial Proteins ◽  
Isolated Mitochondria ◽  
Mitochondrial Inner Membrane ◽  
Carbonyl Cyanide ◽  
A Cell

Ferrochelatase (protohaem ferro-lyase, EC 4.99.1.1), the terminal enzyme of the haem-biosynthetic pathway, is an integral membrane protein of the mitochondrial inner membrane. When murine erythroleukaemia cells are labelled in vivo with [35S]methionine, lysed, and the extract is immunoprecipitated with rabbit anti-(mouse ferrochelatase) antibody, a protein of Mr 40,000 is isolated. However, when isolated mouse RNA is translated in a cell-free reticulocyte extract, a protein of Mr 43,000 is isolated. Incubation of this Mr 43,000 protein with isolated mitochondria resulted in processing of the Mr 43,000 precursor to the Mr 40,000 mature-sized protein. Addition of carbonyl cyanide m-chlorophenylhydrazone and/or phenanthroline inhibits this processing. These data indicate that ferrochelatase, like most mitochondrial proteins, is synthesized in the cytoplasm as a larger precursor and is then translocated and processed to a mature-sized protein in an energy-required step.

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