scholarly journals Combining site-directed spin labeling in vivo and in-cell EPR distance determination

2020 ◽  
Vol 22 (9) ◽  
pp. 4875-4879 ◽  
Author(s):  
Pia Widder ◽  
Julian Schuck ◽  
Daniel Summerer ◽  
Malte Drescher
Keyword(s):  
Amino Acid ◽  
Stop Codon ◽  
Spin Labeling ◽  
Amino Acid Incorporation ◽  
Distance Determination ◽  
E Coli ◽  
Acid Incorporation ◽  
Canonical Amino Acid ◽  
Site Directed Spin Labeling

Non-canonical amino acid incorporation via amber stop codon suppression and in vivo site-directed spin labeling allow in-cell EPR distance determination in E. coli.

Reduced and S-carboxymethylated human growth hormone: a probe for diabetogenic action

1984 ◽  
Vol 247 (5) ◽  
pp. E639-E644
Author(s):  
C. M. Cameron ◽  
J. L. Kostyo ◽  
J. A. Rillema ◽  
S. E. Gennick
Keyword(s):  
Growth Hormone ◽  
Amino Acid ◽  
Human Growth Hormone ◽  
Human Growth ◽  
Mouse Mammary Gland ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Hypophysectomized Rats

The biological activity profile of reduced and S-carboxymethylated human growth hormone (RCM-hGH) was determined to establish its suitability for study of the diabetogenic property of hGH. RCM-hGH was found to have greatly attenuated in vivo growth-promoting activity in the 9-day weight-gain test in hypophysectomized rats (approximately 1%) and to have a similar low order of in vitro activity in stimulating amino acid incorporation into the protein of the isolated rat diaphragm. RCM-hGH also only had approximately 1% of the in vitro insulin-like activity of the native hormone on isolated adipose tissue from hypophysectomized rats. In contrast, RCM-hGH retained substantial in vivo diabetogenic activity in the ob/ob mouse, appearing to have approximately 50% of the activity of the native hormone. RCM-hGH was also found to retain significant, although attenuated (25%), in vitro lactogenic activity when tested for the ability to stimulate amino acid incorporation into a casein-rich protein fraction in mouse mammary gland explants. Because RCM-hGH exhibits a high degree of diabetogenic activity, although lacking significant anabolic or insulin-like activities, it will be useful as a "monovalent" probe for the study of the molecular mechanism of the diabetogenic action of GH.

Effect of Cycloheximide on In Vitro and In Vivo Protein Synthesis by Certain Tissues of Rats and Pigeons with Special Reference to Muscle

1973 ◽  
Vol 51 (12) ◽  
pp. 933-941 ◽  
Author(s):  
Njanoor Narayanan ◽  
Jacob Eapen
Keyword(s):  
Amino Acids ◽  
Body Weight ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Amino Acid Incorporation ◽  
Contrast Administration ◽  
Acid Incorporation ◽  
Tissue Homogenates

The effect of cycloheximide in vitro and in vivo on the incorporation of labelled amino acids into protein by muscles, liver, kidneys, and brain of rats and pigeons was studied. In vitro incorporation of amino acids into protein by muscle microsomes, myofibrils, and myofibrillar ribosomes was not affected by cycloheximide. In contrast, administration of the antibiotic into intact animals at a concentration of 1 mg/kg body weight resulted in considerable inhibition of amino acid incorporation into protein by muscles, liver, kidneys, and brain. This inhibition was observed in all the subcellular fractions of these tissues during a period of 10–40 min after the administration of the precursor. Tissue homogenates derived from in vivo cycloheximide-treated animals did not show significant alteration in in vitro amino acid incorporation with the exception of brain, which showed a small but significant enhancement.

scholarly journals Biotin-mediated protein biosynthesis

1974 ◽  
Vol 140 (3) ◽  
pp. 549-556 ◽  
Author(s):  
R. L. Boeckx ◽  
K. Dakshinamurti
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Rat Liver ◽  
Intestinal Mucosa ◽  
Protein Biosynthesis ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Stimulation Of

The effect of administration of biotin to biotin-deficient rats on protein biosynthesis was studied. Biotin treatment resulted in stimulation by more than twofold of amino acid incorporation into protein, both in vivo and in vitro in rat liver, pancreas, intestinal mucosa and skin. Analysis of the products of amino acid incorporation into liver proteins in vivo and in vitro indicated that the synthesis of some proteins was stimulated more than twofold, but others were not stimulated at all. This indicates a specificity in the stimulation of protein synthesis mediated by biotin.

Incorporation of C14-amino acids into protein of isolated diaphragms: effect of epinephrine and norepinephrine

1960 ◽  
Vol 198 (1) ◽  
pp. 54-56 ◽  
Author(s):  
Ira G. Wool
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Glucose Concentration ◽  
Muscle Protein ◽  
Effective Concentration ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Minimal Effective Concentration

When diaphragms isolated from normal rats were incubated with a C14-amino acid the addition of epinephrine or norepinephrine decreased incorporation of C14 into muscle protein. The inhibition occurred whether epinephrine was added in vitro or administered in vivo. The minimal effective concentration of epinephrine in vitro was 0.1 µg/ml. When the glucose concentration in the medium was raised to 300 mg % or more the epinephrine induced inhibition of amino acid incorporation into muscle protein was no longer observed.

scholarly journals Agranular membranes in free polysome preparations and their possible interference in studies of protein biosynthesis

1969 ◽  
Vol 112 (3) ◽  
pp. 269-274 ◽  
Author(s):  
C. N. Murty ◽  
T. Hallinan
Keyword(s):  
Amino Acid ◽  
Functional Capacity ◽  
Initial Rate ◽  
Protein Biosynthesis ◽  
Amino Acid Incorporation ◽  
Electron Microscopic ◽  
Acid Incorporation ◽  
Microscopic Studies ◽  
Purification Methods

1. Phospholipid-rich membranous contaminants are present in free polysomes from rat liver isolated on discontinuous sucrose gradients. 2. Electron-microscopic studies indicate that the membranous contaminants are mainly agranular with very occasional granular membranes. This is confirmed by the study of their sedimentation behaviour and their initial rate of labelling with radioactive glucosamine in vivo. 3. Conventional ribosome-purification methods fail to remove the contaminants, whereas deoxycholate effectively solubilizes the membranous contaminants with little breakdown of polysomes. 4. Amino acid-incorporation studies show that these membranous contaminants may seriously interfere in assessment of the functional capacity of free polysomes in protein biosynthesis in vivo.

scholarly journals Synthesis of mitochondrial protein in the flight muscles of the Colorado beetle. Differentiation in vivo between extra- and intra-mitochondrial contributions to the amino acid incorporation into mitochondrial protein

1973 ◽  
Vol 136 (3) ◽  
pp. 795-802 ◽  
Author(s):  
A. K. M. Bartelink ◽  
C. A. D. De Kort
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Mitochondrial Protein ◽  
Leucine Incorporation ◽  
Amino Acid Incorporation ◽  
Linear Rate ◽  
Synthesis Conditions ◽  
Acid Incorporation ◽  
Flight Muscles

By using cycloheximide, an inhibitor of cytoplasmic protein synthesis, conditions were investigated to estimate in vivo the extra- and intra-mitochondrial contributions to the synthesis of organelle protein in the flight muscles of Colorado beetles. With 4-day-old beetles about 15% of the [14C]leucine incorporation into mitochondrial protein is resistant to the action of cycloheximide. The incorporation into cytosol protein is inhibited by more than 99.5% with cycloheximide. During the first hour after precursor administration the incorporation into mitochondrial protein proceeds, in both the presence and the absence of cycloheximide, at a more-or-less linear rate with time. The cycloheximide-resistant amino acid incorporation is sensitive to the inhibitor of mitochondrial protein synthesis, chloramphenicol. The uncertainties inherent in the use of cycloheximide were discussed in arriving at the conclusion that about 15% of the mitochondrial protein is formed inside the organelle.

scholarly journals Tracking of Engineered Bacteria In Vivo Using Nonstandard Amino Acid Incorporation

2018 ◽  
Vol 7 (6) ◽  
pp. 1640-1650 ◽  
Author(s):  
Pichet Praveschotinunt ◽  
Noémie-Manuelle Dorval Courchesne ◽  
Ilona den Hartog ◽  
Chaochen Lu ◽  
Jessica J. Kim ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Engineered Bacteria
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