Site Specific Incorporation of Amino Acid Analogues into Proteins In Vivo

2000 ◽  
Author(s):  
Anne K. Kowal ◽  
Caroline Kohrer ◽  
Uttam L. RajBhandary
Keyword(s):  
Amino Acid ◽  
Site Specific ◽  
Amino Acid Analogues

scholarly journals Properties of phosphoenolpyruvate carboxykinase (guanosine triphosphate) synthesized in hepatoma cells in the presence of amino acid analogues

1975 ◽  
Vol 146 (3) ◽  
pp. 585-593 ◽  
Author(s):  
S E Knowles ◽  
J M Gunn ◽  
L Reshef ◽  
R W Hanson ◽  
F J Ballard
Keyword(s):  
Amino Acid ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Hepatoma Cells ◽  
Dibutyryl Cyclic Amp ◽  
Enzyme Degradation ◽  
Amino Acid Analogues ◽  
Enzyme Molecules ◽  
Natural Amino Acids

1. Phosphoenolpyruvate carboxykinase (GTP) was induced by a combination of dibutyryl cyclic AMP, theophyline and dexamethasone in Reuber H35 hepatoma cells under conditions where an amino acid in the medium was replaced by an appropriate analogue. 2. With canavanine replacing arginine or with 5-fluorotryptophan or 6-fluorotryptophan replacing tryptophan the induced enzyme had a lower catalytic activity-relative to antibody reactivity. 3. These aberrant enzyme molecules were heat-labile in vitro. 4. Measurements of enzyme degradation in vivo indicated that the canavanine-containing enzyme and the 6-fluorotryptophan-containing enzyme were degraded more rapidly than the enzyme containing all natural amino acids.

Engineering an acetyllysine reader with photocrosslinking amino acid for interactome profiling

2021 ◽  
Author(s):  
Babu Sudhamalla ◽  
Anirban Roy ◽  
Soumen Barman ◽  
Jyotirmayee Padhan
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Protein Interactions ◽  
Unnatural Amino Acids ◽  
Protein Protein Interactions ◽  
Site Specific ◽  
Powerful Approach

The site-specific installation of light-activable crosslinker unnatural amino acids offers a powerful approach to trap transient protein-protein interactions both in vitro and in vivo. Herein, we engineer a bromodomain to...

Site Specific Combinations of Stabilizing and Destabilizing Amino Acid Replacements in Yeast Cytochrome c: In Vivo and in Vitro Effects

Biochemistry ◽  
1995 ◽  
Vol 34 (21) ◽  
pp. 7103-7112 ◽  
Author(s):  
Lisa I. Linske-O'Connell ◽  
Fred Sherman ◽  
George McLendon
Keyword(s):  
Amino Acid ◽  
Cytochrome C ◽  
Site Specific ◽  
In Vitro Effects

scholarly journals Increased degradation rates of protein synthesized in hepatoma cells in the presence of amino acid analogues

1975 ◽  
Vol 146 (3) ◽  
pp. 595-600 ◽  
Author(s):  
S E Knowles ◽  
J M Gunn ◽  
R W Hanson ◽  
F J Ballard
Keyword(s):  
Amino Acid ◽  
Cell Cultures ◽  
Incubation Medium ◽  
Hepatoma Cells ◽  
Cell Protein ◽  
Chase Period ◽  
Separate Cell ◽  
Degradation Rates ◽  
Amino Acid Analogues

1. Reuber H35 hepatoma cells incorporate the arginine analogue canavanine into cell protein when arginine is omitted from the incubation medium. 2. By labelling arginine-containing proteins with (14-C)leucine and then canavanine-containing proteins with (3-H)leucine in the same cells, it is possible to measure the degradation of both types of protein during a subsequent ‘chase’ period. With this technique it has been shown that canavanine-containing proteins are degraded at a rate severalfold greater than normal proteins. Comparable results were found when 6-fluorotryptophan was used as an analogue to tryptophan. 3. Control experiments in which the labelling order was reversed or where the animo acid and its analogue were incubated in separate cell cultures support the conclusion that abberrant proteins are rapidly degraded in vivo.

scholarly journals KINETICS OF AMINO ACID INCORPORATION INTO SERUM PROTEINS

1955 ◽  
Vol 38 (3) ◽  
pp. 283-293 ◽  
Author(s):  
H. Green ◽  
H. S. Anker
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Serum Protein ◽  
Transit Time ◽  
Serum Proteins ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Amino Acid Analogues ◽  
Kinetics Of

1. The effect of varying body temperature on the rate of amino acid incorporation into serum protein does not give support to the idea that the rate of this process is adjusted in vivo to restore those protein molecules destroyed by thermal denaturation. The experimentally observed Q10 was about 3.9. 2. When amino acids are injected into the blood of animals in a steady state of serum protein turnover, a period of time elapses before these amino acids can be found in the serum proteins. This has been called transit time. At a given temperature (31°) it is the same in rabbits, turtles, and Limulus (1 hour). In rabbits and turtles it has a Q10 of 3.2. It appears to be specifically related to the process of synthesis (or release) of serum proteins. 3. It was not possible to affect the transit time or the incorporation rate by the administration of amino acid analogues.

scholarly journals Genetically Encoded Tetrazine Amino Acid Directs Rapid Site-Specific in Vivo Bioorthogonal Ligation with trans-Cyclooctenes

2012 ◽  
Vol 134 (6) ◽  
pp. 2898-2901 ◽  
Author(s):  
Jason L. Seitchik ◽  
Jennifer C. Peeler ◽  
Michael T. Taylor ◽  
Melissa L. Blackman ◽  
Timothy W. Rhoads ◽  
...  
Keyword(s):  
Amino Acid ◽  
Site Specific

scholarly journals Interaction of the FimB Integrase with thefimS Invertible DNA Element in Escherichia coliIn Vivo and In Vitro

2000 ◽  
Vol 182 (10) ◽  
pp. 2953-2959 ◽  
Author(s):  
Lesley S. Burns ◽  
Stephen G. J. Smith ◽  
Charles J. Dorman
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence Analysis ◽  
Specific Component ◽  
Genetic Switch ◽  
Site Specific ◽  
Tyrosine Recombinases ◽  
Catalytic Motif ◽  
A Site

ABSTRACT The FimB protein is a site-specific recombinase that inverts thefimS genetic switch in Escherichia coli. Based on amino acid sequence analysis alone, FimB has been assigned to the integrase family of tyrosine recombinases. We show that amino acid substitutions at positions R47, H141, R144, and Y176, corresponding to highly conserved members of the catalytic motif of integrase proteins, render FimB incapable of inverting the fimS element in vivo. The arginine substitutions reduced the ability of FimB to bind tofimS in vivo or in vitro, while the substitution R144Q resulted in a protein unable to bind independently to the half sites located at the left end of fimS in phase-on bacteria. These data confirm that FimB is an integrase and suggest that residue R144 has a role in binding to a specific component of the fimswitch.

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