Protein synthesis in artificial cells: using compartmentalisation for spatial organisation in vesicle bioreactors

In recent years, researchers have been pursuing a method to design and to construct life forms from scratch — in other words, to create artificial cells. In many studies, artificial cellular membranes have been successfully fabricated, allowing the research field to grow by leaps and bounds. Moreover, in addition to lipid bilayer membranes, proteins are essential factors required to construct any cellular metabolic reaction; for that reason, different cell-free expression systems under various conditions to achieve the goal of controlling the synthetic cascades of proteins in a confined area have been reported. Thus, in this review, we will discuss recent issues and strategies, enabling to control protein synthesis cascades that are being used, particularly in research on artificial cells.

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Long-lived protein expression in hydrogel particles: towards artificial cells

Chemical Science ◽

10.1039/c8sc00383a ◽

2018 ◽

Vol 9 (18) ◽

pp. 4275-4279 ◽

Cited By ~ 20

Author(s):

Xiaoyu Zhou ◽

Han Wu ◽

Miao Cui ◽

Sze Nga Lai ◽

Bo Zheng

Keyword(s):

Gene Regulation ◽

Protein Expression ◽

Artificial Cells ◽

Hydrogel Particles ◽

Artificial Cell ◽

Genetic Oscillators

A new artificial cell was capable of long-lived protein expression and supported gene regulation and genetic oscillators.

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Combining in Vitro Folding with Cell Free Protein Synthesis for Membrane Protein Expression

Biochemistry ◽

10.1021/acs.biochem.6b00488 ◽

2016 ◽

Vol 55 (30) ◽

pp. 4212-4219 ◽

Cited By ~ 22

Author(s):

Paul J. Focke ◽

Christopher Hein ◽

Beate Hoffmann ◽

Kimberly Matulef ◽

Frank Bernhard ◽

...

Keyword(s):

Protein Synthesis ◽

Membrane Protein ◽

Protein Expression ◽

Free Protein ◽

Membrane Protein Expression ◽

Cell Free Protein Synthesis

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In Vitro Protein Synthesis in Semipermeable Artificial Cells

ACS Synthetic Biology ◽

10.1021/acssynbio.1c00044 ◽

2021 ◽

Author(s):

Damian Van Raad ◽

Thomas Huber

Keyword(s):

Protein Synthesis ◽

Artificial Cells ◽

In Vitro Protein Synthesis ◽

Vitro Protein

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Increased Protein Synthesis by Human Platelets during Phagocytosis of Latex Particles in Vitro

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647928 ◽

1976 ◽

Vol 35 (02) ◽

pp. 350-357 ◽

Cited By ~ 8

Author(s):

Hana Bessler ◽

Galila Agam ◽

Meir Djaldetti

Keyword(s):

Protein Synthesis ◽

Fold Increase ◽

Human Platelets ◽

Polystyrene Latex ◽

Latex Particles ◽

The Third ◽

Platelet Protein ◽

Dose Dependent ◽

Phagocytotic Activity

SummaryA three-fold increase of protein synthesis by human platelets during in vitro phagocytosis of polystyrene latex particles was detected. During the first two hours of incubation, the percentage of phagocytizing platelets and the number of latex particles per platelet increased; by the end of the third hour, the first parameter remained stable, while the number of latex particles per cell had decreased.Vincristine (20 μg/ml of cell suspension) inhibited platelet protein synthesis. This effect was both time- and dose-dependent. The drug also caused a decrease in the number of phagocytizing cells, as well as in their phagocytotic activity.

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EFFECT OF ACTINOMYCIN D ON TSH-INDUCED STIMULATION OF THYROIDAL PROTEIN SYNTHESIS IN THE RAT

Acta Endocrinologica ◽

10.1530/acta.0.0770064 ◽

1974 ◽

Vol 77 (1) ◽

pp. 64-70 ◽

Cited By ~ 9

Author(s):

Gustav Wägar

Keyword(s):

Protein Synthesis ◽

Actinomycin D ◽

The Other ◽

Leucine Incorporation ◽

Short Term ◽

Other Hand ◽

Thyroid Glands ◽

Translational Level ◽

Stimulation Of

ABSTRACT Whether the short-term regulation of thyroidal protein synthesis by TSH occurs at the transcriptional or the translational level was tested by measuring the effect of actinomycin D (act D) on the TSH-induced stimulation of L-14C-leucine incorporation into the thyroidal proteins of rats. TSH was injected 6 h before the rats were killed. The thyroid glands were then removed and incubated in vitro in the presence of L-14C-leucine for 2 h. The pronounced stimulation of leucine incorporation in the TSH-treated animals was depressed as compared with controls but still significant even when the animals had been pre-treated with 100 μg act D 24 and 7 h before sacrifice. On the other hand, act D strongly decreased incorporation of 3H-uridine into RNA. Short-term regulation of thyroidal protein synthesis by TSH appears to be partly but not wholly dependent on neosynthesis of RNA. Hence regulation may partly occur at the translation level of protein synthesis.

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INHIBITION OF THYROGLOBULIN BIOSYNTHESIS AND DEGRADATION BY EXCESS IODIDE. SYNERGISM WITH LITHIUM

Acta Endocrinologica ◽

10.1530/acta.0.0810495 ◽

1976 ◽

Vol 81 (2) ◽

pp. 495-506 ◽

Cited By ~ 9

Author(s):

A. Radvila ◽

R. Roost ◽

H. Bürgi ◽

H. Kohler ◽

H. Studer

Keyword(s):

Protein Synthesis ◽

Thyroid Gland ◽

Inhibitory Action ◽

Inhibit Protein Synthesis ◽

Thyrotrophic Hormone ◽

Thyroid Glands ◽

Pre Treatment ◽

Excess Iodide ◽

Kidney Liver

ABSTRACT Lithium and excess iodide inhibit the release of thyroid hormone from preformed stores. We thus tested the hypothesis that this was due to an inhibition of thyroglobulin breakdown. Rats were pre-treated with propylthiouracil (PTU) for 3 weeks in order to deplete their thyroids of thyroglobulin. While the PTU was continued, lithium chloride (0.25 mEq./100 g weight) or potassium iodide (3 mg per rat) were injected every 12 h for 3 days. Thereafter the thyroglobulin content in thyroid gland homogenates was measured. PTU pre-treatment lowered the thyroglobulin content from 4.21 to 0.22 mg/100 mg gland. Lithium caused a marked re-accumulation of thyroglobulin to 0.60 mg/100 mg within 3 days. While iodide alone had only a borderline effect, it markedly potentiated the action of lithium and a combination of the two drugs increased the thyroglobulin content to 1.04 mg/100 mg. Thyroxine was injected into similarly pre-treated animals to suppress secretion of thyrotrophic hormone. This markedly inhibited the proteolysis of thyroglobulin and 1.3 mg/100 mg gland accumulated after 3 days. Excess iodide, given in addition to thyroxine, decreased the amount of thyroglobulin accumulated to 0.75 mg/100 mg gland. To study whether this could be explained by an inhibitory action of iodide on thyroglobulin biosynthesis, thyroid glands from animals treated with excess iodide were incubated in vitro in the presence of 0.2 mm iodide for 3 h. Iodide decreased the incorporation of radioactive leucine into total thyroidal protein and into thyroglobulin by 25 and 35 % respectively. Iodide did not inhibit protein synthesis in the kidney, liver or muscle tissue. Thus, large doses of iodide selectively inhibit thyroglobulin biosynthesis.

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Long noncoding RNA OIP5-AS1 overexpression promotes viability and inhibits high glucose-induced oxidative stress of cardiomyocytes by targeting microRNA-34a/SIRT1 axis in diabetic cardiomyopathy

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530321666201230090742 ◽

2020 ◽

Vol 21 ◽

Author(s):

Haiyun Sun ◽

Chong Wang ◽

Ying Zhou ◽

Xingbo Cheng

Keyword(s):

Oxidative Stress ◽

Protein Expression ◽

Diabetic Cardiomyopathy ◽

High Glucose ◽

Long Noncoding Rna ◽

Noncoding Rna ◽

Luciferase Reporter ◽

H9c2 Cells ◽

Promoting Effect

Objective: Diabetic cardiomyopathy (DCM) is an important complication of diabetes. This study was attempted to discover the effects of long noncoding RNA OIP5-AS1 (OIP5-AS1) on the viability and oxidative stress of cardiomyocyte in DCM. Methods: The expression of OIP5-AS1 and microRNA-34a (miR-34a) in DCM was detected by qRT-PCR. In vitro, DCM was simulated by high glucose (HG, 30 mM) treatment in H9c2 cells. The viability of HG (30 mM)-treated H9c2 cells was examined by MTT assay. The reactive oxygen species (ROS), superoxide dismutase (SOD) and malondialdehyde (MDA) levels were used to evaluate the oxidative stress of HG (30 mM)-treated H9c2 cells. Dual-luciferase reporter assay was used to confirm the interactions among OIP5-AS1, miR-34a and SIRT1. Western blot was applied to analyze the protein expression of SIRT1. Results: The expression of OIP5-AS1 was down-regulated in DCM, but miR-34a was up-regulated. The functional experiment stated that OIP5-AS1 overexpression increased the viability and SOD level, while decreased the ROS and MDA levels in HG (30 mM)-treated H9c2 cells. The mechanical experiment confirmed that OIP5-AS1 and SIRT1 were both targeted by miR-34a with the complementary binding sites at 3′UTR. MiR-34a overexpression inhibited the protein expression of SIRT1. In the feedback experiments, miR-34a overexpression or SIRT1 inhibition weakened the promoting effect on viability, and mitigated the reduction effect on oxidative stress caused by OIP5-AS1 overexpression in HG (30 mM)-treated H9c2 cells. Conclusions: OIP5-AS1 overexpression enhanced viability and attenuated oxidative stress of cardiomyocyte via regulating miR-34a/SIRT1 axis in DCM, providing a new therapeutic target for DCM.

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